BRPI0621906A2 - DNA molecule encoding a mutant histidine kinase polypeptide and its related use, mutant histidine kinase polypeptide, as well as method of producing a genetically modified plant - Google Patents

Abstract

<B>MOLéCULA DE DNA QUE CODIFICA UM POLIPEPTìDEO DE HISTIDINA QUINASE MUTANTE E SEU USO RELACIONADO, POLIPEPTìDEO DE HISTIDINA QUINASE MUTANTE, BEM COMO MéTODO DE PRODUçãO DE UMA PLANTA GENETICAMENTE MODIFICADA<D>. A presente invenção refere-se à formação de nódulos radiculares fixadores de nitrogênio em leguminosas que é induzida pela captação de moléculas sinalizadoras de oligossacarídeo de lipoquitina secretadas por bactérias Rhizobium compatíveis, que desencadeiam uma via simbiótíca comum. A presente invenção provê um mutante de formação espontânea de nódulo (snf2), no qual a formação de nódulos simbióticos é espontânea, levando ao desenvolvimento nodular na ausência bem como na presença de bactérias Rhizobium e/ou sinais rizobiais exógenos. A invenção ainda provê uma sequência de DNA isolada que codifica uma histidina quinase mutante independente de citocinina, cuja atividade resulta neste fenótipo dominante de 'ganho de função' da formação espontânea de nódulo. Além disso, mostra-se que o gene snf2 confere um fenótipo caracterizado pela organogênese regulada de nódulos espontâneos em plantas que têm um contexto genético de nodulação deficiente. Um gene da invenção, que confere este fenótipo de nodulação espontânea, tem utilidade para transferência e estabelecimento da capacidade de fixação de nitrogênio em plantas não-noduladas, e por meio disso reduz a dependência de fertilizantes nitrogenados das culturas de plantas não-noduladas.<B> DNA MOLECULE THAT CODES A MUTANT HYSTIDINE KINASE POLYPEPTIDE AND RELATED USE, MUTANT HYSTIDINE KINASE POLYPEPTIDE, AS WELL AS A GENETICALLY MODIFIED PLANT PRODUCTION METHOD <D>. The present invention relates to the formation of nitrogen-fixing root nodules in legumes that is induced by the uptake of lipoquitin oligosaccharide signaling molecules secreted by compatible Rhizobium bacteria, which trigger a common symbiotic pathway. The present invention provides a spontaneous nodule formation mutant (snf2), in which the formation of symbiotic nodules is spontaneous, leading to nodular development in the absence as well as in the presence of Rhizobium bacteria and / or exogenous rhizobial signals. The invention further provides an isolated DNA sequence encoding a cytokinin-independent mutant histidine kinase, whose activity results in this dominant 'function gain' phenotype of spontaneous nodule formation. Furthermore, it is shown that the snf2 gene confers a phenotype characterized by the regulated organogenesis of spontaneous nodules in plants that have a defective nodulation genetic context. A gene of the invention, which confers this spontaneous nodulation phenotype, is useful for transferring and establishing the nitrogen fixing capacity in non-nodulated plants, and thereby reduces the nitrogen fertilizer dependence on non-nodulated plant cultures.

Description

Relatório Descritivo da Patente de Invenção para "MOLÉCULADE DNA QUE CODIFICA UM POLIPEPTÍDEO DE HISTIDINA QUINASEMUTANTE E SEU USO RELACIONADO, POLIPEPTÍDEO DE HISTIDINAQUINASE MUTANTE, BEM COMO MÉTODO DE PRODUÇÃO DE UMAPLANTA GENETICAMENTE MODIFICADA".Patent Descriptive Report for "DNA MOLECULADE ENCODING A HYSTIDINE KINASEMUTANT POLYPEPTIDE AND ITS RELATED USE, MUTANT HISTIDINAKINASE POLYPEPTIDE, AS A METHOD OF PRODUCTION OF A GENETICALLY MODIFIED APPLANTA".

Antecedentes da InvençãoBackground of the Invention

O crescimento das culturas agrícolas é geralmente limitado peladisponibilidade de nitrogênio, e pelo menos 50% da demanda global é supri-da pela aplicação de fertilizantes sintéticos na forma de amônia, nitrato ouuréia. Contudo, há uma necessidade crescente de explorar uma das fontesnaturais de nitrogênio mais importantes para agricultura, ou seja, a fixaçãobiológica de nitrogênio.Crop growth is generally limited by the availability of nitrogen, and at least 50% of global demand is supplied by the application of synthetic fertilizers in the form of ammonia, nitrate or urea. However, there is a growing need to explore one of the most important natural nitrogen sources for agriculture, namely biological nitrogen fixation.

A fonte primária de fixação biológica de nitrogênio são Rhizobi-um ou Rhizobia spp e a actinobactéria Frankia spp, que são um pequenogrupo de procariotos que produz nitrogenases e forma associações endos-simbióticas com plantas conferindo a capacidade de fixar nitrogênio. Emboramuitas plantas possam associar-se com bactérias fixadoras de nitrogênio,somente algumas plantas, todas membros do Ciado Rosídea I, formam umaassociação endossimbiótica com a Rhizobia spp e Frankia spp, que são asúnicas em que a maior parte do nitrogênio é transferida e assimilada pelaplanta hospedeira. As plantas da família Leguminosae, que inclui soja, feijão,ervilha, amendoim, grão-de-bico, feijão-caupi, lentilha, guandu, alfafa e trevo,são os membros agronomicamente mais importantes deste pequeno grupode plantas fixadoras de nitrogênio. A fixação biológica de nitrogênio atravésda associação endossimbiótica reduz a necessidade de fertilizantes nitroge-nados caros na cultura de Ieguminosas e é uma importante característica daagricultura sustentável. As Ieguminosas também podem utilizar o nitrogêniodisponível no solo, tal que quando os níveis de nitrato do solo são altos, aformação de nódulo é suprimida e a planta substitui o metabolismo de nitro-gênio por crescimento em nitrato (Wopereis et ai, 2000).The primary source of biological nitrogen fixation is Rhizobi-um or Rhizobia spp and actinobacteria Frankia spp, which are a small group of prokaryotes that produce nitrogen and form endosymbiotic associations with plants that confer the ability to fix nitrogen. Although many plants may associate with nitrogen-fixing bacteria, only a few plants, all members of Rosicea Ciado, form an endosymbiotic association with Rhizobia spp and Frankia spp, which are the only ones in which most nitrogen is transferred and assimilated by the host plant. . Plants in the Leguminosae family, which include soybeans, beans, peas, peanuts, chickpeas, cowpeas, lentils, guandu, alfalfa, and clover, are the most agronomically important members of this small group of nitrogen-fixing plants. Biological nitrogen fixation through endosymbiotic association reduces the need for expensive nitrogenous fertilizers in leguminous crops and is an important feature of sustainable agriculture. Ieguminous can also use nitrogen available in the soil, such that when soil nitrate levels are high, nodule formation is suppressed and the plant replaces nitrate metabolism with nitrate growth (Wopereis et al, 2000).

A simbiose Rhizobium-\egurr\\nosa inclui a interação de uma sé-rie de genes de planta e bacterianos em um processo complexo que leva àiniciação e desenvolvimento de nódulos radiculares. A organogênese dosnódulos é desencadeada pelo microssimbionte rizobial, mas a planta Iegu-minosa hospedeira codifica o programa de desenvolvimento responsável porconstruir os tecidos do nódulo e por regular o processo. O baixo peso mole-cular dos oligossacarídeos de Iipoquitina (fatores Nod), sintetizados e secre-tados pelo Rizóbio, são as principais moléculas sinalizadoras que desenca-deiam este processo. O principal fator Nod secretado pelo Mesorhizobiumloti, microssimbionte do Lotus, é uma N-acetilglicosamina pentamérica queporta um ácido cis-vacênico e um grupo carbamoíla no resíduo terminal nãoreduzido junto a uma 4-O-acetilfucose no resíduo terminal reduzido. A cap-tação do fator Nod no Lotus é mediada pelas quinases receptoras NFR1 eNFR5 (Radutoiu et ai., 2003 Nature 425: 585-592; Madsen et ai., 2003 Natu-re 425: 637-640), que em conjunto com um gene da quinase receptora LRR,SymRK, comunicam-se com uma via comum de transdução de sinal, com-partilhada com a simbiose micorrizal (Oldroyd e Downie, 2004 Moi. Celi Bio-Iogy 5: 566-576). Essa via comum é codificada por sete genes, SymRK, Cas-tor, Pollux, Nup133, CCaMK [Sym15], Sym6 e Sym24. A análise de mutantestem mostrado que receptor (es) NFR1/NFR5, SymRK que codifica a proteínaquinase LRR, canal(is) de cátion CASTOR/POLLUX e nucleoporina133 sãonecessários para a variação de cálcio, uma das mais precoces respostas fisio-lógicas detectáveis nos pelos radiculares expostos ao fator Nod purificado.Rhizobium-egurr \\ nosa symbiosis includes the interaction of a series of plant and bacterial genes in a complex process that leads to the initiation and development of root nodules. The organogenesis of the nodules is triggered by the rhizobial microsymbionte, but the host Iegminosa plant encodes the developmental program responsible for constructing the nodule tissues and regulating the process. The low molecular weight of Ipoquitin oligosaccharides (Nod factors), synthesized and secreted by the Rhizobium, are the main signaling molecules that trigger this process. The main Nod factor secreted by Mesorhizobiumloti, the microsymbiont of the Lotus, is a pentameric N-acetylglycosamine carrying a cis-vaccenic acid and a carbamoyl group in the unreduced terminal residue along with a 4-O-acetylfucose in the reduced terminal residue. Nod factor uptake in Lotus is mediated by NFR1 and NFR5 receptor kinases (Radutoiu et al., 2003 Nature 425: 585-592; Madsen et al., 2003 Natu-re 425: 637-640), which together with an LRR receptor kinase gene, SymRK, communicate with a common signal transduction pathway, shared with mycorrhizal symbiosis (Oldroyd and Downie, 2004 Moi. Celi Bio-Iogy 5: 566-576). This common pathway is encoded by seven genes, SymRK, Castor, Pollux, Nup133, CCaMK [Sym15], Sym6 and Sym24. Mutant analysis has shown that NFR1 / NFR5 receptor (s), SymRK encoding LRR protein kinase, CASTOR / POLLUX cation channel (s) and nucleoporin133 are required for calcium variation, one of the earliest physiological responses detectable in hair. roots exposed to purified Nod factor.

Para estabelecer a simbiose, o Rizóbio tem acesso a células in-dividuais da planta por endocitose onde são instalados em simbiossomasrodeados por uma membrana peribacterioide. Em Lotus, a infecção ocorreatravés de um filamento de infecção que leva as bactérias através dos pelosradiculares no córtex radicular e as distribui às células, que se tornam infec-tadas com o simbiossoma que contém células fixadoras de nitrogênio. Emresposta às bactérias introduzidas, os pelos radiculares se deformam e enro-lam, montando uma pequena bolsa que provê um sítio para recepção dofilamento de infecção (Geurts et ai, 2005 Curr. Opinion Plant Biol., 8: 346-352). Os filamentos de infecção são estruturas derivadas das plantas, origi-nados da invaginação da membrana plasmática, acompanhada pela deposi-ção externa de material da parede celular. Frente ao progresso interno dofilamento intracelular, células corticais radiculares desdiferenciam e reentramno ciclo celular para iniciar o primórdio de nódulo. Mais tarde, no processo, aformação padrão e a diferenciação celular especificam tipos de tecido e célu-Ia inclusive as células infectadas que endocitaram os Rizóbios. No nódulomaduro funcional, os feixes vasculares periféricos estão conectados à vas-culatura radicular e os principais tipos de tecidos/célula podem ser distingui-dos (Pawlowski e Bisseling, 1996, Plant Cell 8: 1899-1913).To establish symbiosis, Rhizobium has access to individual plant cells by endocytosis where they are installed in symbiosomes surrounded by a peribacterioid membrane. In Lotus, infection occurs through a strand of infection that carries bacteria through the root hairs in the root cortex and distributes them to cells, which become infected with the symbiotome containing nitrogen-fixing cells. In response to the introduced bacteria, the root hairs deform and coil, assembling a small pouch that provides a site for receiving dofilament of infection (Geurts et al., 2005 Curr. Opinion Plant Biol., 8: 346-352). Infection filaments are plant-derived structures that originate from plasma membrane invagination, accompanied by the external deposition of cell wall material. Faced with internal progress in intracellular thickening, root cortical cells de-differentiate and reenter the cell cycle to initiate the nodule primordium. Later in the process, standard deformation and cell differentiation specify tissue types and cells including the infected cells that endocyted Rhizobia. In the functional nodule, the peripheral vascular bundles are connected to the root vasculature and the main tissue / cell types can be distinguished (Pawlowski and Bisseling, 1996, Plant Cell 8: 1899-1913).

A análise de um grupo de mutantes de nodulação, inclusive al-guns que falham em mostrar oscilações de cálcio em resposta aos sinais dofator Nod, revelou que além da falta de nodulação, esses mutantes são in-capazes de formar endossimbioses com fungos micorrizais arbusculares.Isso implica que uma via comum de transdução de sinal simbiótica é com-partilhada por dois tipos de relações endossimbióticas, ou seja, simbiose denódulo radicular, que é basicamente restrita à família das leguminosas, efungos micorrizais arbusculares, que é comum à maioria das espécies deplantas terrestres. Isto sugere que possa haver alguns genes-chave dosquais as leguminosas dispõem para empregar na nodulação, e que estãofaltando em cultura de plantas tais como cereais. A identificação desses ge-nes-chave, que codificam funções que são indispensáveis para o estabele-cimento de um sistema de fixação de nitrogênio em leguminosas, e suatransferência e expressão em plantas não-noduladas, tem sido por muitotempo um objetivo de melhoradores moleculares de plantas. Isto poderia terum impacto agronômico significante no cultivo de cereais como arroz, onde aprodução de duas colheitas por ano pode necessitar de fertilização com até400 kg de nitrogênio por hectare.Analysis of a group of nodulation mutants, including some that fail to show calcium oscillations in response to Nod dofactor signals, revealed that in addition to lack of nodulation, these mutants are unable to form endosymbioses with arbuscular mycorrhizal fungi. This implies that a common pathway of symbiotic signal transduction is shared by two types of endosymbiotic relationships, namely root-symbiosis symbiosis, which is basically restricted to the legume family, arbuscular mycorrhizal fungi, which is common to most plant species. terrestrial. This suggests that there may be some key genes that legumes have to employ in nodulation, and that they are lacking in plant culture such as cereals. The identification of these key genes, which encode functions that are indispensable for the establishment of a nitrogen fixation system in legumes, and their transfer and expression in non-nodulated plants, has long been a goal of molecular enhancers. plants. This could have a significant agronomic impact on cereal crops such as rice, where the production of two harvests per year may require fertilization with up to 400 kg of nitrogen per hectare.

A simbiose de nódulo radicular depende de uma interação bem-sucedida entre a planta hospedeira e o seu simbionte cognato que inclui aetapa do reconhecimento do fator nod pela planta hospedeira. A identifica-ção de genes que regulam a nodulação em Leguminosas forneceria ferra-mentas para otimizar e modificar este processo em benefício da agricultura.Root node symbiosis depends on a successful interaction between the host plant and its cognate symbiote that includes the step in recognizing the nodule factor by the host plant. Identifying genes that regulate nodulation in legumes would provide tools to optimize and modify this process for the benefit of agriculture.

Em resumo, há uma necessidade de melhorar a capacidade deformação de nódulos e as propriedades de fixação de nitrogênio nas culturasde leguminosas, bem como transferir essa via para a cultura não-noduladapara suprir as necessidades nutricionais de uma população global crescente,minimizando o uso futuro de fertilizantes nitrogenados e o seu impacto am-biental negativo associado.In summary, there is a need to improve nodule deformation capacity and nitrogen-fixing properties in leguminous crops, as well as to transfer this pathway to non-nodular crops to meet the nutritional needs of a growing global population while minimizing the future use of legumes. nitrogen fertilizers and their associated negative environmental impact.

Sumário da InvençãoSummary of the Invention

Uma primeira modalidade da invenção é uma molécula de DNAque codifica um polipeptídeo de histidina quinase mutante compreendendouma seqüência de aminoácidos selecionada entre: SEQ ID NO: 6, 9, 12, 14-20, em que o resíduo de aminoácido correspondente a Xaa é selecionadoentre isoleucina, serina, treonina, valina, metionina, alanina, fenilalanina, ti-rosina, triptofano, arginina, lisina, glicina, histidina, aspartato, asparagina,glutamato, glutamina, prolina e cisteína,A first embodiment of the invention is a DNA molecule encoding a mutant histidine kinase polypeptide comprising an amino acid sequence selected from: SEQ ID NO: 6, 9, 12, 14-20, wherein the amino acid residue corresponding to Xaa is selected from isoleucine , serine, threonine, valine, methionine, alanine, phenylalanine, thi-rosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine, proline and cysteine,

um ortólogo de (a) e um truncamento de (a) ou (b), capaz de in-duzir formação espontânea de nódulo em uma planta.an ortholog of (a) and a truncation of (a) or (b), capable of inducing spontaneous nodule formation in a plant.

Uma segunda modalidade da invenção é um polipeptídeo dehistidina quinase mutante composto de uma seqüência de aminoácidos se-lecionada entre: (a) SEQ ID NO: 6, 9, 12, 14-20, em que o resíduo de ami-noácido correspondente a Xaa é selecionado entre isoleucina, serina, treoni-na, valina, metionina, alanina, fenilalanina, tirosina, triptofano, arginina, lisi-na, glicina, histidina, aspartato, asparagina, glutamato, glutamina, prolina ecisteína; (b) uma variante alélica de (a) independente de citocinina; (c) umortólogo de (a) independente de citocinina; e (d) um truncamento de (a), (b)ou (c).A second embodiment of the invention is a mutant dehistidine kinase polypeptide composed of an amino acid sequence selected from: (a) SEQ ID NO: 6, 9, 12, 14-20, wherein the amino acid residue corresponding to Xaa is selected from isoleucine, serine, threone, valine, methionine, alanine, phenylalanine, tyrosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine, proline and cysteine; (b) an allelic variant of (a) cytokine independent; (c) a cytokine independent (a )ortologist; and (d) a truncation of (a), (b) or (c).

Uma modalidade adicional da invenção é uma planta genetica-mente modificada caracterizada por ter uma seqüência de nucleotídeos quecodifica um polipeptídeo compreendendo um mutante de histidina quinaseconsistindo em uma seqüência de aminoácidos selecionada entre: (a) SEQID NO: 6, 9, 12, 14-20, em que o resíduo de aminoácido correspondente aXaa é selecionado entre isoleucina, serina, treonina, valina, metionina, alani-na, fenilalanina, tirosina, triptofano, arginina, lisina, glicina, histidina, asparta-to, asparagina, glutamato, glutamina, prolina e cisteína; (b) uma variante alé-lica de (a) independente de citocinina; (c) um ortólogo de (a) independentede citocinina; e (d) um truncamento de (a), (b) ou (c), em que a dita planta écapaz da formação espontânea de nódulo.A further embodiment of the invention is a genetically modified plant characterized in that it has a nucleotide sequence that encodes a polypeptide comprising a histidine kinase mutant consisting of an amino acid sequence selected from: (a) SEQID NO: 6, 9, 12, 14- 20, wherein the amino acid residue corresponding to AXaa is selected from isoleucine, serine, threonine, valine, methionine, alanine, phenylalanine, tyrosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine proline and cysteine; (b) an allelic variant of (a) cytokine independent; (c) an ortholog of (a) cytokine independent; and (d) a truncation of (a), (b) or (c), wherein said plant is capable of spontaneous nodule formation.

A invenção é ainda direcionada ao uso de uma molécula de áci-do nucleico que codifica uma histidina quinase mutante consistindo em umaseqüência de aminoácidos selecionada entre: (a) SEQ ID NO: 7, 8, 9, 10, 11,15, 26 e 27, em que o resíduo de aminoácido correspondente a Xaa é sele-cionado entre isoleucina, serina, treonina, valina, metionina, alanina, fenila-lanina, tirosina, triptofano, arginina, lisina, glicina, histidina, aspartato, aspa-ragina, glutamato, glutamina, prolina e cisteína; (b) um ortólogo de (a); e (c)um truncamento de (a) ou (b), como um transgene para produzir a plantageneticamente modificada da invenção, capaz de nodulação espontânea deacordo com suas várias modalidades.The invention is further directed to the use of a nucleic acid molecule encoding a mutant histidine kinase consisting of an amino acid sequence selected from: (a) SEQ ID NO: 7, 8, 9, 10, 11,15, 26 and 27, wherein the amino acid residue corresponding to Xaa is selected from isoleucine, serine, threonine, valine, methionine, alanine, phenyl lanine, tyrosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine, proline and cysteine; (b) an ortholog of (a); and (c) a truncation of (a) or (b) as a transgene to produce the plantagenetically modified invention capable of spontaneous nodulation according to its various embodiments.

A invenção é ainda direcionada a um método de produção deuma planta geneticamente modificada de acordo com a invenção nas suasvárias modalidades, caracterizada pela introdução de um cassete de genecompreendendo a dita seqüência de nucleotídeos que codifica o dito poli-peptídeo e selecionando uma planta transgênica e sua progênie que expres-sa o dito polipeptídeo.The invention is further directed to a method of producing a genetically modified plant according to the invention in its various embodiments, characterized by the introduction of a genet cassette comprising said nucleotide sequence encoding said polypeptide and selecting a transgenic plant and its progeny expressing said polypeptide.

A invenção ainda inclui uma planta geneticamente modificadaproduzida de acordo com um processo de mutagênese de DNA e seleção deuma planta capaz de formação espontânea de nódulo, ou por um método detransformação com um transgene que codifica uma histidina quinase mutan-te da invenção.The invention further includes a genetically modified plant produced according to a DNA mutagenesis process and selection of a plant capable of spontaneous nodule formation, or by a transgene method encoding a mutant histidine kinase of the invention.

A invenção ainda inclui uma semente ou uma cultura obtida daplanta geneticamente modificada da invenção. Além disso, a invenção é di-recionada ao uso de uma planta geneticamente modificada de acordo com ainvenção em um programa de melhoramento, e uma planta selecionada noprograma de melhoramento compreendendo uma seqüência de nucleotídeosque codifica um polipeptídeo compreendendo uma histidina quinase mutanteconsistindo em uma seqüência de aminoácidos selecionada entre: (a) SEQID NO: 6, 9, 12, 14-20 em que o resíduo de aminoácido correspondente aXaa é selecionado entre isoleucina, serina, treonina, valina, metionina, alani-na, fenilalanina, tirosina, triptofano, arginina, lisina, glicina, histidina, asparta-to, asparagina, glutamato, glutamina, prolina e cisteína; (b) uma variante alé-lica de (a) independente de citocinina; (c) um ortólogo de (a) independentede citocinina, e (d) um truncamento de (a), (b) ou (c), em que a dita planta écapaz de formação espontânea de nódulo.The invention further includes a seed or culture obtained from the genetically modified plant of the invention. In addition, the invention is directed to the use of a genetically modified plant according to the invention in a breeding program, and a plant selected in the breeding program comprising a nucleotide sequence encoding a polypeptide comprising a mutant histidine kinase consisting of a mutant histidine kinase sequence. amino acids selected from: (a) SEQID NO: 6, 9, 12, 14-20 wherein the amino acid residue corresponding to AXaa is selected from isoleucine, serine, threonine, valine, methionine, alani-na, phenylalanine, tyrosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine, proline and cysteine; (b) an allelic variant of (a) cytokine independent; (c) an ortholog of (a) cytokinin independentede, and (d) a truncation of (a), (b) or (c), wherein said plant is capable of spontaneous nodule formation.

Breve Descrição dos DesenhosBrief Description of the Drawings

Figura 1. Caracterização fenotípica do mutante snf2.Figure 1. Phenotypic characterization of snf2 mutant.

(A) rizóbios do tipo selvagem induziram o nódulo radicular (B)nódulo radicular espontâneo na raiz snf2 5 semanas após a germinação. Ascabeças de seta indicam nódulos. Secção transversal do (C) tipo selvagem e(D) da raiz snf2 no tempo zero e (E) tipo selvagem e (F) raiz snf2 após 6 diasde crescimento no meio B5 sem hormônio. A cabeça de seta indica célulasse dividindo no periciclo. As setas indicam células do xilema. (G) e (H) cres-cimento do calo a partir de hipocótilos do tipo selvagem e snf2 em diferentesconcentrações de auxina e citocinina. Os calos foram fotografados depois de21 dias de crescimento a 26°C. (I) e (J) segmentos radiculares do tipo selva-gem e snf2 incubados por três semanas em meio sem hormônio. Barras deescala: (C); (D); (E); (F) 50 pm.(A) wild-type rhizobia induced root nodule (B) spontaneous root nodule in snf2 root 5 weeks after germination. Arrowheads indicate nodules. Cross section of (C) wild type and (D) snf2 root at time zero and (E) wild type and (F) snf2 root after 6 days of growth in B5 medium without hormone. The arrowhead indicates cell dividing on the pericycle. Arrows indicate xylem cells. (G) and (H) callus growth from wild type and snf2 hypocotyls at different concentrations of auxin and cytokine. Corns were photographed after 21 days of growth at 26 ° C. (I) and (J) jungle-like and snf2 root segments incubated for three weeks in hormone-free medium. Scaling Bars: (C); (D); (AND); (F) 50 pm.

Figura 2. Clonagem baseada no mapa do gene LhklFigure 2. Cloning based on Lhkl gene map

A: Figura do cromossomo IV de Lotus japonicus com mapa ge-nético estendido da região abaixo do snf2 (Iocus Lhk1), compreendendo 6clones de BAC/TAC (sublinhado em negrito). As linhas verticais indicam mi-crossatélites ou marcadores de polimorfismo de nucleotídeo único. O núme-ro das plantas recombinantes obtidas por um marcador de microssatélite(TM) é indicado em negrito abaixo do marcador.A: Figure of chromosome IV of Lotus japonicus with extended genetic map of the region below snf2 (Iocus Lhk1), comprising 6 BAC / TAC clones (underlined in bold). Vertical lines indicate micromolatites or single nucleotide polymorphism markers. The number of recombinant plants obtained by a microsatellite (TM) marker is indicated in bold below the marker.

Β: A estrutura éxon-íntron do gene Lhk1, onde os 11 éxons deLhkl são indicados por caixas abertas. A transição CDT no éxon 4 do alelomutante snf2 (indicado com uma cabeça de seta preta) codifica um polipep-tídeo mutante LHK1 que compreende uma substituição de F266 para L266.Β: The exon-intron structure of the Lhk1 gene, where the 11 exons of Lhkl are indicated by open boxes. The CDT transition in exon 4 of the snf2 allelomutant (indicated with a black arrowhead) encodes a mutant LHK1 polypeptide comprising a replacement of F266 to L266.

Figura 3. Transformação do alelo snf2 no tipo selvagem nas raí-zes cabeludas leva à organogênese controlada e fenótipo de nodulação es-pòntânea.Figure 3. Wild-type transformation of the snf2 allele in the hairy roots leads to controlled organogenesis and spontaneous nodulation phenotype.

Plantas Lotus japonicus do tipo selvagem foram transformadascom Agrobacterium rhizogenes portando o gene snf2. Nódulos espontâneosnas raízes cabeludas estão identificados por cabeças de seta brancas. A raizprincipal está identificada por uma seta branca.Wild-type Lotus japonicus plants were transformed with Agrobacterium rhizogenes carrying the snf2 gene. Spontaneous nodules in the hairy roots are identified by white arrowheads. The main root is identified by a white arrow.

Figura 4. Estrutura da proteína LHK1 de Lotus.Figure 4. Structure of Lotus LHK1 protein.

(A) Representação esquemática dos domínios da proteínaLHK1. (B) A seqüência de aminoácidos de LHK1 arranjada em domínios pro-teicos. O domínio extracelular do receptor predito é dado em itálico e o do-mínio CHASE dentro do domínio extracelular do receptor está sublinhado; odomínio histidina quinase é dado em negrito e sublinhado; o domínio ATPa-se His Quinase é dado em negrito; e o domínio do receptor está em negrito eitálico. O asterisco no domínio CHASE marca a posição da substituição doaminoácido no alelo snf2.(A) Schematic representation of LHK1 protein domains. (B) The amino acid sequence of LHK1 arranged in protein domains. The extracellular domain of the predicted receptor is given in italics and the domain CHASE within the extracellular domain of the receptor is underlined; The histidine kinase domain is bold and underlined; the ATPa domain and His Kinase is given in bold; and the receptor domain is in bold italic. The asterisk in the CHASE domain marks the amino acid substitution position in the snf2 allele.

Figura 5. Ensaios in vivo de sinalização de citocinina mediadapelo receptor.Figure 5. In vivo receptor mediated cytokine signaling assays.

(A) Ensaio em placa da atividade de β-galactosidase expressapor um gene repórter cpsr.iacZ em E. coli. Uma cepa de E. coli SRC122 queporta o repórter de cpsr.iacZ, transformada com um construto de plasmídeocompreendendo o DNAc de snf2 ou o DNAc do tipo selvagem, foi cultivadaem placas na ausência de citocinina ou em presença de quatro citocininasdiferentes. A cor azul mostra a conversão do substrato X-Gal pela β-galactosidase.(A) Plaque assay for β-galactosidase activity by expressing a cpsr.iacZ reporter gene in E. coli. An E. coli SRC122 strain carrying the cpsr.iacZ reporter, transformed with a plasmid construct comprising snf2 cDNA or wild type cDNA, was cultured in plaques in the absence of cytokine or in the presence of four different cytokines. The blue color shows the conversion of X-Gal substrate by β-galactosidase.

(B) Indução da atividade de β-galactosidase pela citocinina emculturas líquidas de SRC122 cpsr.iacZ transformada com o DNAc de snf2 oucom DNAc do tipo selvagem. T-z: Trans-zeatina.(B) Induction of β-galactosidase activity by cytokine in SRC122 cpsr.iacZ liquid cultures transformed with snf2 cDNA or wild type cDNA. T-z: Transzeatin.

(C) Modelo de trabalho do papel funcional de Lhkl na nodula-ção. O reconhecimento de um fator Nod rizobial corretamente adornado peloNFR1 e NFR5 induz transdução de sinal através da via comum, incluindovariação de cálcio e a interpretação da variação de cálcio pela proteínaCCaMK. Um aumento localizado na biossíntese de citocinina captado peloreceptor LHK1 leva então à desdiferenciação celular e ativação do ciclo celu-lar. snf2 está constitutivamente ativo mas ainda necessita dos genes Nin eSym35 para a organogênese do nódulo.(C) Working model of Lhkl's functional role in nodulation. Recognition of a correctly adorned rhizobial Nod factor by NFR1 and NFR5 induces signal transduction through the common pathway, including calcium variation and the interpretation of calcium variation by the CCaMK protein. A localized increase in cytokine biosynthesis captured by LHK1 receptor then leads to cell de-differentiation and activation of the cell cycle. snf2 is constitutively active but still needs the Nin eSym35 genes for nodule organogenesis.

Figura 6. Efeito do tratamento com fito-hormônio no crescimentode calo em explantes de raiz.Figure 6. Effect of phyto-hormone treatment on callus growth in root explants.

O crescimento de calo no ecotipo Gifu do L. japonicus do tiposelvagem (A) e (B) fragmentos de raiz snf2 em diferentes concentrações deauxina e citocinina. Os calos foram fotografados após 21 dias de crescimen-to a 26°C.Callus growth in the Gifu ecotype of wild type L. japonicus (A) and (B) snf2 root fragments at different concentrations of deauxin and cytokine. Corns were photographed after 21 days of growth at 26 ° C.

Figure 7. Expressão do gene Lhkl em órgãos e Lhk1, Lrr5 e ninem resposta à citocinina.Figure 7. Lhkl gene expression in organs and Lhk1, Lrr5 and ninem response to cytokine.

(A) Expressão de Lhkl em órgãos diferentes. (B) Expressão dogene Lrr5 em explantes de raiz do tipo selvagem e snf2 incubados em placade ágar-ágar B5 com ou sem 0,5 pg/ml de BAP durante 10 dias. (C), (D), (E)Expressão dos genes Lrr5, Lhkl e Nin, em raízes intactas do tipo selvageme snf2 em resposta à citocinina. (F) Expressão do gene Nin em explantes deraiz do tipo selvagem e snf2 incubados em placas de ágar-ágar B5 com ousem 0,5 pg/ml de BAP durante 10 dias.(A) Lhkl expression in different organs. (B) Lrr5 dogene expression in wild type and snf2 root explants incubated on B5 agar agar with or without 0.5 pg / ml BAP for 10 days. (C), (D), (E) Expression of Lrr5, Lhkl and Nin genes in intact wild-type and snf2 roots in response to cytokine. (F) Expression of the Nin gene in wild-type and snf2 deerizal explants incubated on B5 agar plates with dare 0.5 pg / ml BAP for 10 days.

Figura 8: Efeito da citocinina no comprimento do broto e da raizde L. japonicus do tipo selvagem e no mutante snf2-2.Figure 8: Effect of cytokine on wild-type L. japonicus shoot and root length and snf2-2 mutant.

Plantas foram cultivadas em % de B&D contendo diferentes con-centrações de BAP. O comprimento do broto e da raiz foi medido após 3 se-manas de crescimento. (A) comprimento do broto (B) comprimento da raiz.Plants were grown in% B&D containing different BAP concentrations. Sprout and root length were measured after 3 weeks of growth. (A) bud length (B) root length.

Figure 9. Fenótipo de nodulação de snf2 e do duplo mutantesnf2 har1 na ausência dos rizóbios.Figure 9. Snf2 and double mutant nf2 har1 nodulation phenotype in the absence of rhizobia.

(A) snf2 mutante e (B) duplo mutante snf2 har1 5 semanas apósa germinação.(A) snf2 mutant and (B) double mutant snf2 har1 5 weeks after germination.

Descrição Detalhada da InvençãoDetailed Description of the Invention

A presente invenção provê um gene isolado que codifica um po-lipeptídeo mutante, cuja expressão em plantas confere um fenótipo de nodu-lação espontânea. Mais especificamente o gene foi isolado de mutantes denodulação espontânea snf2 do Lotus japonicus que desenvolvem nódulossem rizóbios brancos na ausência do microssimbionte M. loti, enquanto asplantas Lotus do tipo selvagem somente são noduladas depois da induçãocom os seus cognatos fatores nod ou simbionte rizobial (figura 1a, b). Análi-ses histológicas detalhadas de secções de nódulos demonstram que os nó-dulos espontâneos snf2 são nódulos genuínos com uma ontogenia e fisiolo-gia semelhantes aos nódulos rizobiais induzidos. Assim, os nódulos radicula-res espontâneos formados em mutantes snf2 originam-se de células corti-cais e têm uma vasculatura periférica característica de nódulos do tipo sel-vagem, embora eles sejam desprovidos de filamentos de infecção e rizóbios.O alelo snf2 é monogênico dominante e a inoculação de mutantes snf2 como M. Ioti resulta no desenvolvimento de nódulos radiculares normais fixado-res de nitrogênio compatível com a presença de uma mutação de ganho defunção neste alelo.The present invention provides an isolated gene encoding a mutant po-lipeptide whose expression in plants confers a spontaneous nodulation phenotype. More specifically, the gene has been isolated from spontaneous snf2 mutation mutations in the Lotus japonicus that develop nodules without white rhizobia in the absence of M. loti microstrips, while wild-type Lotus plants are only nodulated after induction with their cognate nodular or rhizobial symbiotic factors (Figure 1a , B). Detailed histological analyzes of nodule sections show that spontaneous snf2 nodules are genuine nodules with similar ontogeny and physiology as induced rhizobial nodules. Thus, spontaneous root nodules formed in snf2 mutants originate from cortical cells and have a peripheral vasculature characteristic of wild-type nodules, although they are devoid of infection filaments and rhizobia. The snf2 allele is monogenic and the inoculation of snf2 mutants such as M. Ioti results in the development of normal nitrogen-fixing root nodules compatible with the presence of a defective gain mutation in this allele.

O alelo snf2 localiza-se no cromossomo 4 em uma posição a-proximadamente a 1 cM do fim do braço longo (figura 2). O gene, corres-pondente ao alelo snf2, codifica uma forma mutante de uma Lotus histidinaquinase (LHK1) que é um homólogo dos genes Arabidopsis histidina quinase(AHK) que codificam as proteínas receptoras de citocinina. Uma comparaçãodo gene Lotus histidina quinase do tipo selvagem (LhkI) [SEQ ID NO:1] como gene correspondente do alelo snf2 [SEQ ID NO:4] revela uma transição denucleotídeo único (C para T) na seqüência de codificação do alelo snf2 queresulta na substituição de uma Ieucina conservada [L266 em Lhk1\ SEQ IDNO:3] por uma fenilalanina (F266 no alelo snf2 do Lhk1\ SEQ ID NO:6) nopolipeptídeo codificado. A invenção ainda provê DNAcs isolados correspon-dentes ao transcrito do gene Lhkl do tipo selvagem [SEQ ID NO:2] e ao ale-lo snf2 [SEQ ID NO:5]. O alinhamento das seqüências genômicas do Lhkl eseqüências de DNAc ou o respectivo gene do alelo snf2 e seqüências deDNAc, define uma estrutura primária de Lhkl e o seu alelo snf2 como com-posto de 11 éxons (figura 2).The snf2 allele is located on chromosome 4 in a position approximately 1 cM from the end of the long arm (Figure 2). The gene, corresponding to the snf2 allele, encodes a mutant form of a Lotus histidine kinase (LHK1) that is a homologue to the Arabidopsis histidine kinase (AHK) genes that encode cytokine receptor proteins. A comparison of the wild-type Lotus histidine kinase (LhkI) gene [SEQ ID NO: 1] as the corresponding snf2 allele gene [SEQ ID NO: 4] reveals a single nucleotide transition (C to T) in the snf2 allele coding sequence and result replacing a conserved eukine [L266 in Lhk1 \ SEQ IDNO: 3] with a phenylalanine (F266 in the snh2 allele of Lhk1 \ SEQ ID NO: 6) encoded polypeptide. The invention further provides isolated cDNAs corresponding to the wild type Lhkl gene transcript [SEQ ID NO: 2] and to the snf2 ale [SEQ ID NO: 5]. Alignment of Lhkl genomic sequences and cDNA sequences or the respective snf2 allele gene and DNANA sequences defines a primary structure of Lhkl and its snf2 allele as a composite of 11 exons (Figure 2).

A invenção ainda provê a proteína receptora de citocinina(LHK1) isolada, do tipo selvagem e mutante, codificada, respectivamente,pelo gene Lhkl e seu alelo snf, compreendendo 993 aminoácidos com umamassa predita de 110 kD (figura 4). Ambas as proteínas compreendem doissegmentos transmembrana no N-terminal, localizados entre os aminoácidos37 e 57 e entre os aminoácidos 328 e 357. Localizado entre esses segmen-tos estão os motivos característicos de ciclases/histidina quinases associa-das ao domínio sensorial extracelular (CHASE). Este domínio extracelularpredito é seguido por uma histidina quinase intracelular putativa entre os a -minoácidos 379 e 693 e um domínio receptor entre os aminoácidos 852 e985. Esses domínios são característicos de sistemas reguladores de doiscomponentes que funcionam por uma fosforrealocação. A fenilalanina 266no alelo snf2, substituída pela Ieucina 266, está localizada em um domínioconservado compartilhado entre os domínios extracelulares CHASE de re-ceptores histidina quinase (figura 4). As propriedades funcionais da histidinaquinase (proteína receptora de citocinina) codificada pelo alelo snf2 incluema atividade independente de citocinina, tal que o alelo snf2, em contrastecom alelo do tipo selvagem Lhkl pode induzir a nodulação na ausência desinalização de citocinina.The invention further provides the isolated wild-type and mutant cytokine receptor protein (LHK1) encoded by the Lhk1 gene and its snf allele, respectively, comprising 993 amino acids with a predicted mass of 110 kD (Figure 4). Both proteins comprise N-terminal transmembrane disruptions, located between amino acids37 and 57 and between amino acids 328 and 357. Located between these segments are the characteristic motifs of cyclases / histidine kinases associated with extracellular sensory domain (CHASE). . This predicted extracellular domain is followed by a putative intracellular histidine kinase between amino acids 379 and 693 and a receptor domain between amino acids 852 and 985. These domains are characteristic of two-component regulatory systems that function by phosphorealocation. Phenylalanine 266 in the snf2 allele, replaced by Ieucine 266, is located in a conserved domain shared between the extracellular CHASE domains of histidine kinase receptors (Figure 4). The functional properties of histidine kinase (cytokine receptor protein) encoded by the snf2 allele includes cytokine independent activity, such that the snf2 allele, in contrast to wild type Lhkl allele, can induce nodulation in the absence of cytokine de-signaling.

Assim sendo, a presente invenção provê uma proteína histidinaquinase mutante isolada (proteína mutante receptora de citocinina) capaz deinduzir a nodulação espontânea quando expressa em uma planta, em que oresíduo de aminoácido correspondente a L266 na histidina quinase LHK1 dotipo selvagem codificado pelo gene Lhkl do Lotus japonicus é substituídopor um aminoácido selecionado entre isoleucina, serina, treonina, valina,metionina, alanina, fenilalanina, tirosina, triptofano, arginina, lisina, glicina,histidina, aspartato, asparagina, glutamato, glutamina, prolina e cisteína. Al-ternativamente, o resíduo de aminoácido correspondente à L266 na histidinaquinase LHK1 do tipo selvagem (Xaa) é conservativamente substituído comum aminoácido selecionado entre fenilalanina, tirosina, triptofano, histidinaou prolina. O dito último grupo pode ser aumentado para incluir qualquer dos3 grupos seguintes de aminoácidos (arginina, lisina, aspartato, glutamato) ou(asparagina, glutamina, serina, treonina) ou (cisteína, metionina, isoleucina,valina, glicina, alanina). Essa histidina quinase mutante da invenção incluihomólogos de várias plantas nos quais o resíduo de aminoácido correspon-dente a L266 na histidina quinase LHK1 do tipo selvagem (com base no ali-nhamento como mostrado na Tabela 2) é deletado ou substituído com qual-quer outro aminoácido diferente de leucina. Mais especificamente, a inven-ção inclui os homólogos de histidina quinase mutante do Lotus japonicus(forma mutante de Lhk1; SEQ ID NO:6,15), M. truncatula (forma mutante deABE94286; SEQ ID N0:16), Arabidopsis (forma mutante de BAB33311; SEQID NO:9,17), arroz (forma mutante de XP_469566; SEQ ID N0:18), milho(forma mutante de BAE80688; SEQ ID NO:12,19) e Cucurbita maxima (for-ma mutante de CAF31355.1; O SEQ ID N0:14,20) ou ortólogos ou variantesalélicas das mesmas, em que Xaa é qualquer outro aminoácido diferente deleucina. Em uma modalidade, a histidina quinase mutante é aquela na qualXaa é selecionada a partir do grupo da fenilalanina, tirosina, triptofano, histi-dina ou prolina, ou alternativamente Xaa é fenilalanina. O ortólogo ou varian-te alélica da histidina quinase mutante da invenção compartilha uma porcen-tagem de identidade com a seqüência da dita histidina quinase mutante, se-lecionada do grupo composto de pelo menos 60, 65, 70, 75, 80, 85, 90, 95, e98 por cento, ou truncamento desses, em que a dita quinase tem todas aspropriedades funcionais da histidina quinase mutante da invenção, tal quequando expressa em uma planta da invenção ela confere a capacidade deformar nódulos espontâneos. Em uma modalidade adicional o alelo ortólo-go/variante é uma histidina quinase mutante que é uma proteína receptorade citocinina que é independente citocinina.Accordingly, the present invention provides an isolated mutant histidine kinase protein (cytokine receptor mutant protein) capable of inducing spontaneous nodulation when expressed in a plant, wherein the amino acid residue corresponding to L266 in the wild type LHK1 histidine kinase encoded by the Lotus Lhkl gene japonicus is replaced by an amino acid selected from isoleucine, serine, threonine, valine, methionine, alanine, phenylalanine, tyrosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine, proline and cysteine. Alternatively, the amino acid residue corresponding to L266 in wild-type LHK1 histidine kinase (Xaa) is conservatively substituted with the amino acid selected from phenylalanine, tyrosine, tryptophan, histidine or proline. Said latter group may be increased to include any of the following 3 amino acid groups (arginine, lysine, aspartate, glutamate) or (asparagine, glutamine, serine, threonine) or (cysteine, methionine, isoleucine, valine, glycine, alanine). Such mutant histidine kinase of the invention includes various plant homologs in which the amino acid residue corresponding to L266 in the wild-type histidine kinase LHK1 (based on alignment as shown in Table 2) is deleted or replaced with any other amino acid other than leucine. More specifically, the invention includes mutant histidine kinase homologues of Lotus japonicus (mutant form of Lhk1; SEQ ID NO: 6.15), M. truncatula (mutant form ofABE94286; SEQ ID NO: 16), Arabidopsis (mutant form BAB33311 mutant; SEQID NO: 9.17), rice (XP_469566 mutant form; SEQ ID NO: 18), maize (BAE80688 mutant form; SEQ ID NO: 12.19) and Cucurbita maxima (mutant form of CAF31355.1; SEQ ID NO: 14,20) or orthologs or allelic variants thereof, wherein Xaa is any amino acid other than deleucine. In one embodiment, the mutant histidine kinase is one in which Xaa is selected from the group of phenylalanine, tyrosine, tryptophan, histidine or proline, or alternatively Xaa is phenylalanine. The ortholog or allelic variant of the mutant histidine kinase of the invention shares an identity percentage with the sequence of said mutant histidine kinase, selected from the group consisting of at least 60, 65, 70, 75, 80, 85, 90, 95, and 98 percent, or truncation thereof, wherein said kinase has all the functional properties of the inventive mutant histidine kinase, such that when expressed in a plant of the invention it confers the ability to deform spontaneous nodules. In an additional embodiment the ortho-go / variant allele is a mutant histidine kinase which is a cytokine receptor protein that is cytokine independent.

Esta substituição única de aminoácido ou deleção codificadapelo alelo mutante snf2 do gene Lhk1 ou o seu homólogo é suficiente paraconferir um fenótipo de nodulação espontânea nas raízes das plantas trans-formadas do tipo selvagem que expressam um transgene compreendendo ogene correspondente ao alelo snf mutante (Tabela 1 e figura 3). O alelo snf2atua como um gene dominante de ganho de função, visto que a nodulaçãoespontânea é vista em um contexto genético do tipo selvagem, e está au-sente em plantas transformadas com gene Lhk1 do tipo selvagem. Portanto,a expressão de uma única cópia da histidina quinase mutante da invençãoem uma planta, como no caso de uma planta heterozigota é suficiente paraconferir o fenótipo snf2 de nodulação espontânea.A presente invenção ainda provê uma molécula de DNA que co-difica a dita histidina quinase mutante homóloga (em que Xaa é Phe) do Lo-tus japonicus (SEQ ID NO:4,5), M. truncatula (forma mutante da AcessãoAC141922.19; SEQ ID NO:7), Arabidopsis (forma mutante de AB049935.1;SEQ ID NO:8), arroz (forma mutante de NT_079916.2; SEQ ID NO: 10), mi-lho (forma mutante de AB206392.1; SEQ ID NO:11) e Cucurbita maxima(forma mutante de AJ628045.1; SEQ ID NO:13), ou fragmentos desses quecodificam uma histidina quinase mutante funcional inteira ou truncada capazde causar nodulação espontânea quando expressa em uma planta.This unique amino acid substitution or deletion encoded by the mutant snf2 allele of the Lhk1 gene or its counterpart is sufficient to confer a spontaneous nodulation phenotype on the roots of wild-type transformed transgene plants that express the mutant snf allele gene (Table 1 and figure 3). The snf2 allele acts as a dominant function gain gene, since spontaneous nodulation is seen in a wild type genetic context, and is present in plants transformed with wild type Lhk1 gene. Therefore, the expression of a single copy of the mutant histidine kinase of the invention in a plant, as in the case of a heterozygous plant, is sufficient to confer the spontaneously nodulating snf2 phenotype. The present invention further provides a DNA molecule that co-differs said histidine. homologous mutant kinase (where Xaa is Phe) from Lo-tus japonicus (SEQ ID NO: 4.5), M. truncatula (Accession mutant form AC141922.19; SEQ ID NO: 7), Arabidopsis (AB049935 mutant form. 1; SEQ ID NO: 8), rice (NT_079916.2 mutant form; SEQ ID NO: 10), corn (AB206392.1 mutant form; SEQ ID NO: 11) and Cucurbita maxima (AJ628045 mutant form .1; SEQ ID NO: 13), or fragments thereof which encode an entire or truncated functional mutant histidine kinase capable of causing spontaneous nodulation when expressed in a plant.

De acordo com a presente invenção, a seqüência de DNA quecodifica a proteína histidina quinase mutante da invenção está operavelmen-te ligada a uma seqüência de DNA promotora capaz de dirigir a expressãoda dita histidina quinase em uma planta, e a uma seqüência terminadora 3'.O promotor pode ser um promotor que dirige a expressão da dita histidinaquinase em tecidos radiculares de uma planta e/ou em células destinadas atornar-se o primórdio do nódulo e desenvolver-se em nódulos. Os exemplosconvenientes de um promotor e um terminador incluem o promotor e a se-qüência terminadora correspondente ao gene Lhkl do tipo selvagem. Em umamodalidade da invenção, o promotor usado para dirigir a expressão da histidi-k·· 20 na quinase mutante é um regulado (por exemplo, promotor de tecido ou espe-cífico do tipo celular) que inclui o promotor nativo do gene Lhkl ou o seu ho-mólogo como definido na presente invenção. Um exemplo de um promotorconstitutivo heterólogo inclui o promotor do 35SCaMV (Acc. No: V00141,J02048). Um transgene (cassete de gene) compreendendo uma seqüência deDNA que codifica uma histidina quinase mutante da invenção operavelmentefundida a uma seqüência promotora e opcionalmente a uma seqüência termi-nadora pode ser construído por técnicas de DNA recombinante.In accordance with the present invention, the DNA sequence that encodes the mutant histidine kinase protein of the invention is operably linked to a promoter DNA sequence capable of directing said histidine kinase expression in a plant, and to a 3 'terminator sequence. The promoter may be a promoter that directs expression of said histidine kinase in root tissues of a plant and / or cells designed to bind the nodule primordium and develop into nodules. Suitable examples of a promoter and terminator include the promoter and the terminator sequence corresponding to the wild-type Lhkl gene. In one embodiment of the invention, the promoter used to direct expression of histidi-k ·· 20 in the mutant kinase is a regulated (e.g., tissue-specific or cell-type promoter) that includes the native Lhkl gene promoter or the its homologue as defined in the present invention. An example of a heterologous conductive promoter includes the 35SCaMV promoter (Acc. No: V00141, J02048). A transgene (gene cassette) comprising a DNA sequence encoding a mutant histidine kinase of the invention operably fused to a promoter sequence and optionally to a terminator sequence may be constructed by recombinant DNA techniques.

De acordo com a presente invenção, um transgene compreen-dendo uma seqüência de DNA que codifica a histidina quinase mutante podeser usado para gerar uma planta que expressa a histidina quinase mutanteda invenção. O transgene pode estar integrado de maneira estável no ge-noma de uma planta hospedeira por técnicas de transformação bem-conhecidas pelo versado na técnica. Além disso, os vetores binários e méto-dos baseados em Agrobacterium tumefaciens para a integração estável dostransgenes em todas as principais plantas de cereal são conhecidos, comodescrito por exemplo para arroz (Hiei et al., 1994, The Plant J. 6; 271-282), emilho (Yuji et al., 1996, Nature Biotech. 14: 745-750). Uma seqüência deDNA que codifica uma histidina quinase mutante também pode ser introduzi-da em outra planta pelo cruzamento com uma planta geneticamente modifi-cada que expressa a histidina quinase mutante da invenção.According to the present invention, a transgene comprising a DNA sequence encoding the mutant histidine kinase may be used to generate a plant expressing the mutant histidine kinase of the invention. The transgene may be stably integrated into the bud of a host plant by transformation techniques well known to one skilled in the art. In addition, binary vectors and methods based on Agrobacterium tumefaciens for the stable integration of transgenes into all major cereal plants are known, as described for rice (Hiei et al., 1994, The Plant J. 6; 271- 282), Emilho (Yuji et al., 1996, Nature Biotech. 14: 745-750). A DNA sequence encoding a mutant histidine kinase may also be introduced into another plant by crossing with a genetically modified plant expressing the mutant histidine kinase of the invention.

A planta geneticamente modificada da invenção, se gerada pormutagênese, transformação com um transgene da invenção, ou introduçãodo dito transgene, pode ser usada em um programa de melhoramento, paraselecionar as plantas com a capacidade de fixar nitrogênio, ou capacidadede fixação de nitrogênio aumentada, que herdaram o gene que codifica ahistidina quinase mutante. A invenção assim inclui uma planta geneticamen-te modificada, produzida pela transformação de uma planta natural, que écapaz de nodulação espontânea. A expressão de um gene que codifica umahistidina quinase mutante em uma planta fixadora de nitrogênio, como ummembro da Leguminoseae (tal como soja, feijão, ervilha, amendoim, grão-de-bico, feijão-caupi, lentilha, guandu, alfafa e trevo), tem utilidade particu-larmente com respeito ao aumento da capacidade fixadora de nitrogênio dadita planta sob uma ou mais condições ambientais de crescimento. A ex-pressão de um gene que codifica uma histidina quinase mutante em umacultura que não fixa naturalmente o nitrogênio, como uma planta dicotiledô-nea ou uma planta monocotiledônea inclusive um membro dos cereais (co-mo trigo, centeio, aveia, cevada, sorgo, painço, milho, grama Poaceae e ar-roz), tem particularmente a utilidade com respeito à concessão de habilidadede fixar nitrogênio. As plantas, bem como a progênie da planta, selecionadasem tal programa de melhoramento podem ser cultivadas com o objetivo dacolheita de uma cultura, onde o cultivo pode ser as partes vegetativas daplanta, por exemplo folha, tronco ou tubérculo, ou partes reprodutivas, inclu-sive flores, semente, grãos, espiga ou fruto.The genetically modified plant of the invention, whether generated by mutagenesis, transformation with a transgene of the invention, or introduction of said transgene, may be used in an breeding program to select plants with the ability to fix nitrogen, or increased nitrogen fixation capacity, which inherited the gene encoding mutant ahistidine kinase. The invention thus includes a genetically modified plant produced by the transformation of a natural plant capable of spontaneous nodulation. Expression of a gene encoding a mutant histidine kinase in a nitrogen-fixing plant, such as a Leguminoseae member (such as soy, beans, peas, peanuts, chickpeas, cowpeas, lentils, guandu, alfalfa, and clover) , has particular utility with respect to increasing nitrogen-fixing capacity of each plant under one or more environmental growing conditions. Expression of a gene that encodes a mutant histidine kinase in a non-naturally nitrogen-fixing crop, such as a dicotyledonous or monocotyledonous plant including a member of cereals (such as wheat, rye, oats, barley, sorghum) , millet, maize, grass Poaceae and ar-roz), is particularly useful with respect to granting nitrogen fixation ability. Plants, as well as the progeny of the plant, selected in such a breeding program may be grown for the purpose of harvesting a crop, where the crop may be the vegetative parts of the plant, for example leaf, trunk or tuber, or reproductive parts, including sive flowers, seeds, grains, ear or fruit.

Os exemplos dados abaixo servem para ilustrar várias modali-dades da invenção e as suas respectivas características. Eles demonstramque uma planta, por exemplo o Lotus, que é homozigoto ou heterozigoto pa-ra um gene que codifica o polipeptídeo de histidina quinase mutante da in-venção, forma nódulos espontâneos que podem ser infectados por bactériassimbióticas fixadoras de nitrogênio, por exemplo, o M. loti, e que são capa-zes de fixar o nitrogênio e dar suporte ao crescimento da planta sob condi-ções com nitrogênio limitado. A formação de nódulos fixadores de nitrogêniodepois da inoculação com bactérias fixadoras de nitrogênio na dita plantanodulada espontaneamente não é dependente da produção do fator nod pe-las bactérias infectantes ou captação do fator nod pela planta infectada. Issoindica que somente um subconjunto de genes relacionados à nodulação énecessário para a fixação de nitrogênio ocorrer em um planta nodulada es-pontaneamente que expressa uma histidina quinase mutante depois da ino-culação com uma bactéria rizóbio. As propriedades únicas da histidina qui-nase mutante da invenção podem ser exploradas para aumentar a fixaçãode nitrogênio nas plantas fixadoras de nitrogênio existentes, bem como noestabelecimento de fixação de nitrogênio em plantas não-noduladas.Exemplo 1. Clonagem posicionai e identificação do gene mutante snf-2 emLotus japonicusThe examples given below serve to illustrate various embodiments of the invention and their respective characteristics. They demonstrate that a plant, for example the Lotus, which is homozygous or heterozygous for a gene encoding the invention's mutant histidine kinase polypeptide, forms spontaneous nodules that can be infected with nitrogen-fixing symbiotic bacteria, for example M. loti, which are capable of fixing nitrogen and supporting plant growth under conditions with limited nitrogen. The formation of nitrogen-fixing nodules after inoculation with nitrogen-fixing bacteria in said spontaneously planted nodules is not dependent on the production of the nod factor by the infecting bacteria or uptake of the nod factor by the infected plant. This indicates that only a subset of nodulation-related genes is required for nitrogen fixation to occur in a spontaneously nodulated plant that expresses a mutant histidine kinase after inoculation with a rhizobium bacterium. The unique properties of the inventive mutant histidine kinase can be explored to increase nitrogen fixation in existing nitrogen-fixing plants as well as in establishing nitrogen fixation in non-nodulated plants.Example 1. Positional cloning and identification of the snf mutant gene -2 on Lotus japonicus

Mutantes de Lotus (snf2-1 e snf2-2) têm um fenótipo de nodula-ção espontânea, originado de uma triagem das sementes de Lotus japonicusdo ecotipo Gifu com EMS. O gene mutante snf-2 em Lotus japonicus, que dáorigem à nodulação espontânea, foi localizado no braço longo do cromosso-mo IV, a aproximadamente 1 cM do final, em um Iocus denominado Lhk1.Lotus mutants (snf2-1 and snf2-2) have a spontaneous nodulation phenotype, originated from a screening of Eif Gifu ecotype Lotus japonicus seeds. The mutant snf-2 gene in Lotus japonicus, which gives rise to spontaneous nodulation, was located in the long arm of chromosome IV, approximately 1 cM from the end, in an Iocus called Lhk1.

A posição do gene snf2 foi determinada pelo mapeamento finousando marcadores de microssatélite (marcadores TM) e marcadores poli-mórficos de nucleotídeo único desenvolvidos em clones BAC ou TAC anco-rados ao mapa genético da região Lhk1. O mapeamento foi realizado emuma população F2 estabelecida de um híbrido dos ecotipos de Lotus japoni-eus, Miyakojima e MG 20. O mapa fino foi usado para construir um "contig"físico de TAC/BAC compreendendo seis clones de BAC/TAC de MG20 queforam reunidos para cobrir a região Lhk1 flanqueando entre os dois marca-dores TM1146 e TM0069 (Figure 2). Uma vez que snf2 é uma mutação do-minante, plantas F2 do tipo selvagem, e não as plantas mutantes, forammarcadas para mapeamento dos alvos. A genotipagem de 853 plantas dotipo selvagem que não desenvolveram nódulos radiculares espontaneamen-te foi usada para identificar marcadores que delimitam o Iocus corresponden-te à mutação snf2 a uma região de 120 kb. Quinze genes foram preditosdentro desta região, um dos quais foi predito para codificar um receptor decitocinina. A seqüência dos alelos tanto do tipo selvagem [SEQ ID NO:1]quanto do mutante snf2 [SEQ ID NO:4] deste gene candidato foi determina-da, a qual revelou uma substituição do nucleotídeo C para T em ambos osalelos snf2. O gene de tipo selvagem foi denominado gene histidina quinasede Lotus (LhkI) que codifica um polipeptídeo LHK1 [SEQ ID NO:3]. A transi-ção de nucleotídeo único (C para T) em uma seqüência do éxon do alelomutante snf2 codifica um polipeptídeo que tem uma fenilalanina (F266 emSEQ ID NO:6) em substituição da Ieucina conservada (L266 em SEQ IDNO:3) no polipeptídeo codificado pelo gene de tipo selvagem, e identificasnf2 como um alelo de um gene histidina quinase de Lotus (Lhk1).Exemplo 2. Clonagem e identificação do DNAc de Lhkl correspondente aotranscrito de tipo selvagem do gene Lhkl em Lotus japonicusThe position of the snf2 gene was determined by mapping using microsatellite markers (TM markers) and single nucleotide polymorphic markers developed in BAC or TAC clones anchored to the Lhk1 region genetic map. Mapping was performed on an established F2 population of a hybrid of the Lotus japoni-eus, Miyakojima and MG 20 ecotypes. The thin map was used to construct a physical TAC / BAC contig comprising six MG20 BAC / TAC clones that were joined to cover the Lhk1 region flanking between the two markers TM1146 and TM0069 (Figure 2). Since snf2 is a dominant mutation, wild-type F2 plants, not mutant plants, were marked for target mapping. Genotyping of 853 wild type plants that did not spontaneously develop root nodules was used to identify markers that delimit the Iocus corresponding to the snf2 mutation to a 120 kb region. Fifteen genes were predicted within this region, one of which was predicted to encode a decytokine receptor. The sequence of both wild type [SEQ ID NO: 1] and snf2 mutant [SEQ ID NO: 4] alleles of this candidate gene was determined, which revealed a substitution of nucleotide C for T in both snf2 alleles. The wild-type gene was called the Lotus histidine kinase (LhkI) gene encoding an LHK1 polypeptide [SEQ ID NO: 3]. The single nucleotide (C to T) transition in a snf2 allelomutant exon sequence encodes a polypeptide that has a phenylalanine (F266 in SEQ ID NO: 6) in place of the conserved yeucine (L266 in SEQ IDNO: 3) in the polypeptide encoded by the wild-type gene, and identify nf2 as an allele of a Lotus histidine kinase (Lhk1) gene. Example 2. Cloning and identification of the Lhkl cDNA corresponding to the wild-type Lhkl gene transcript in Lotus japonicus

Um DNAc inteiro de Lhkl (3568 pb) foi isolado de uma biblioteca□ ZAPII de DNAc preparada a partir de RNAm isolado de M. Ioti inoculadanas raízes de Lotus japonicus. O DNAc de Lhkl foi sequenciado [SEQ IDNO:2], no qual o sítio de início de transcrição foi determinado estando situa-do a pelo menos 137 nucleotídeos a montante do códon de iniciação e a se-quência de codificação foi seguida por uma região 3' não traduzida de apro-ximadamente 445 nucleotídeos. O alinhamento das seqüências genômicas ede DNAc definiu uma estrutura primária de Lhkl composta de 11 éxons (Fi-gura 2B). A seqüência de nucleotídeos do alelo mutante snf2 do DNAc doLhkl é indicada SEQ ID NO:5.An entire Lhkl cDNA (3568 bp) was isolated from a cDNA □ ZAPII library prepared from mRNA isolated from M. Ioti inoculated into the roots of Lotus japonicus. Lhkl cDNA was sequenced [SEQ IDNO: 2], in which the transcription initiation site was determined to be at least 137 nucleotides upstream of the initiation codon and the coding sequence was followed by a region 3 'untranslated to approximately 445 nucleotides. Alignment of genomic sequences and cDNA defined a primary Lhkl structure composed of 11 exons (Figure 2B). The nucleotide sequence of the mutant snf2 allele of dohhl cDNA is indicated SEQ ID NO: 5.

Exemplo 3: Expressão do alelo snf2 em raízes de Lotus de tipo selvagemconfere um fenótipo de nodulação espontânea.Example 3: Expression of the snf2 allele in wild-type Lotus roots confers a spontaneous nodulation phenotype.

A expressão transgênica do alelo snf2 de tipo selvagem ou mu-tante em raízes de Lotus foi realizada para confirmar in planta a base genéti-ca do fenótipo de nodulação espontânea. Um fragmento genômico de 12,7kb do clone de BAC 1K18 foi usado para clonar o gene Lhkl de tipo selva-gem incluindo uma região promotora de 2,2 kb, e o seu alelo Snf2 compre-endendo a transição CGT, que então foram subclonados no plasmídeoplV10. Os construtos foram integrados em A. rhizogenes da cepa AR12 pelocruzamento triparental. A transformação foi realizada como descrito porStougaard (1995) Method Mol Biology 49: Plant Gene Transfer and Expres-sion Protocols, p. 49-63, e incluída a transformação com um vetor de contro-le vazio. Nodulação radicular, registrada na ausência da indução rizobial, sófoi detectada em raízes transformadas com o alelo snf2, enquanto o geneLhkl de tipo selvagem falhou em conferir a nodulação espontânea em raízesde tipo selvagem (Tabela 1, figura 3).Transgenic expression of the wild type or mutant snf2 allele in Lotus roots was performed to confirm in plant the genetic basis of the spontaneous nodulation phenotype. A 12.7kb genomic fragment of the BAC 1K18 clone was used to clone the jungle-type Lhkl gene including a 2.2 kb promoter region, and its Snf2 allele comprising the CGT transition, which were then subcloned. in plasmid plV10. The constructs were integrated into A. rhizogenes of the AR12 strain by triparental crossbreeding. Transformation was performed as described by Stouard (1995) Method Mol Biology 49: Plant Gene Transfer and Expression Protocols, p. 49-63, and include transformation with an empty control vector. Root nodule, recorded in the absence of rhizobial induction, was only detected in roots transformed with the snf2 allele, while the wild-type Lhkl gene failed to confer spontaneous nodulation in wild-type roots (Table 1, Figure 3).

Tabela 1Table 1

Indução de nódulos espontâneos em raízes cabeludas de tipo selvagemtransformadas com o gene Lhkl de tipo selvagem ou seu alelode ganho de função snf2Induction of spontaneous nodules in wild-type hairy roots transformed with the wild-type Lhkl gene or its allele of snf2 function gain

<table>table see original document page 17</column></row><table><table> table see original document page 17 </column> </row> <table>

* Raízes transgênicas foram registradas na ausência de M. loti.* Transgenic roots were recorded in the absence of M. loti.

A capacidade do alelo snf2 conferir nodulação espontânea emraízes transformadas que têm o contexto genético de tipo selvagem servepara confirmar a natureza dominante do alelo snf2. Além disso, é surpreen-dentemente mostrado que a histidina quinase independente de citocinina(snf2), quando sob controle de um promotor regulado, por exemplo, promo-tor do gene Lhk1, confere a organogênese controlada de nódulos radicula-res, em contraste com a proliferação celular descontrolada levando ao de-senvolvimento de nódulos volumosos. A formação controlada de nódulosespontâneos, capazes de infecção e fixação de nitrogênio em plantas trans-formadas com o alelo snf2, aumenta sua capacidade fixadora de nitrogêniodurante o cultivo. Uma vez que a única diferença estrutural entre o geneLhkl de tipo selvagem e o alelo snf2 está situada em uma mutação pontualque codifica uma substituição única de aminoácido, esses dados demons-tram que a nodulação espontânea é o resultado de uma substituição L/F266em um polipeptídeo LHK1.The ability of the snf2 allele to confer spontaneous nodulation on transformed roots that have the wild-type genetic context serves to confirm the dominant nature of the snf2 allele. Furthermore, it is surprisingly shown that cytokine-independent histidine kinase (snf2), when under the control of a regulated promoter, for example, the Lhk1 gene promoter, confers the controlled organogenesis of root nodules, in contrast to uncontrolled cell proliferation leading to the development of large nodules. Controlled formation of spontaneous nodules, capable of infection and nitrogen fixation in plants transformed with the snf2 allele, increases their nitrogen fixing capacity during cultivation. Since the only structural difference between wild-type gene Lhkl and snf2 allele is located in a point mutation that encodes a single amino acid substitution, these data show that spontaneous nodulation is the result of an L / F266 substitution on a polypeptide. LHK1.

Exemplo 3: Lotus LHK1 é um membro da família de receptor de citocinina.Example 3: Lotus LHK1 is a member of the cytokine receptor family.

A observação do clone de DNAc do Lhkl revela uma fase aber-ta de leitura de 2979 nucleotídeos que é predita para codificar uma proteínareceptora de citocinina (LHK1) consistindo em 993 aminoácidos com umamassa predita de 110 kD (figura 4). No término N1 dois segmentos trans-membrana estão localizados entre os aminoácidos 37 e 57 e entre os ami-noácidos 328 e 357. Localizado entre esses segmentos estão os motivoscaracterísticos das ciclases/histidina quinases associadas ao domínio senso-rial extracelular (CHASE). Este domínio extracelular predito é seguido poruma histidina quinase intracelular putativa entre os aminoácidos 379 e 693 eum domínio receptor entre os aminoácidos 852 e 985. Esses domínios sãocaracterísticos de sistemas reguladores de dois componentes que funcionampor uma fosforrealocação. A análise comparativa define a proteína Lotus LHK1 como ummembro da família de receptor de citocinina que inclui proteínas de Medica-go truncatula, Arabidopsis, arroz e milho (tabela 2). A proteína Lotus LHK1compartilha uma identidade de seqüência de aminoácidos de 83%, 68%,58% e 49%, respectivamente, com as proteínas homólogas de M. truncatula(ABE94286), Arabidopsis (BAB33311), arroz (XP_469566) e milho(BAD01584). Dentre os três receptores de citocinina Arabidopsis, LHK1 é omais semelhante a AHK4/(Cre1) que é importante para o desenvolvimentoradicular normal e apresenta uma função na captação da citocinina fornecidaexternamente (Mahonen et ai. (2000), Genes Dev. 4: 2938). A Ieucina 266substituída por uma fenilalanina 266 no alelo snf2 está localizada em ummotivo conservado compartilhado entre os domínios CHASE extracelularesdos receptores de histidina quinase (figura 4).Tabela 2Observation of the Lhkl cDNA clone reveals an open reading frame of 2979 nucleotides that is predicted to encode a cytokine protein receptor (LHK1) consisting of 993 amino acids with a predicted 110 kD mass (Figure 4). At the N1 terminus two transmembrane segments are located between amino acids 37 and 57 and between amino acids 328 and 357. Located between these segments are the characteristic motifs of cyclases / histidine kinases associated with extracellular sensory domain (CHASE). This predicted extracellular domain is followed by a putative intracellular histidine kinase between amino acids 379 and 693 and a receptor domain between amino acids 852 and 985. These domains are characteristic of two-component regulatory systems that function by phosphorealocation. Comparative analysis defines the Lotus LHK1 protein as a member of the cytokine receptor family that includes Medica-go truncatula, Arabidopsis, rice and corn proteins (Table 2). Lotus LHK1 protein shares an amino acid sequence identity of 83%, 68%, 58% and 49%, respectively, with the homologous proteins of M. truncatula (ABE94286), Arabidopsis (BAB33311), rice (XP_469566) and corn (BAD01584). ). Of the three Arabidopsis cytokine receptors, LHK1 is the most similar to AHK4 / (Cre1) which is important for normal adult development and plays a role in externally supplied cytokine uptake (Mahonen et al. (2000), Genes Dev. 4: 2938). . Ieucine 266 replaced with a phenylalanine 266 in the snf2 allele is located in a conserved motif shared between the extracellular CHASE domains of the histidine kinase receptors (Figure 4).

<table>table see original document page 19</column></row><table>Tabela 2<table> table see original document page 19 </column> </row> <table> Table 2

<table>table see original document page 20</column></row><table>Tabela 2 - continuacao<table> table see original document page 20 </column> </row> <table> Table 2 - continued

<table>table see original document page 21</column></row><table>Exemplo 4. Atividade independente de Citocinina<table> table see original document page 21 </column> </row> <table> Example 4. Cytokine Independent Activity

O fenótipo de nodulação espontânea conferido pelo alelo snf2 émostrado ser devido a uma função independente de citocinina do receptorde citocinina LHK1 mutante codificado por este alelo, que é caracterizadopor uma substituição L/F266 no domínio CHASE (figura 4). A atividade invivo dos receptores de citocinina Lotus LHK1 de tipo selvagem e mutante e asua resposta à citocinina foi demonstrada por meio de um ensaio de fosfor-realocação de dois componentes desenvolvido em E. coli (Suzuki et ai,(2001) Plant Cell Physioi 42: 107). Neste ensaio a indução de citocinina po-de ser determinada sem interferência de níveis de citocinina endógena, pe-netração de citocinina e vias metabólicas interferentes presentes em plantas.A fosforrealocação explorada neste ensaio está normalmente envolvida naregulação da síntese de polissacarídeo extracelular bacteriano através daativação do operon cps em resposta a um sensor (RcsC)1 o qual tem umaestrutura de domínio semelhante aos receptores de citocinina de planta. Aexpressão funcional de um receptor de citocinina em uma cepa de E. coli desensor RcsC deletado permite a captação de citocinina ser lida como ativi-dade de β-galactosidase pela a expressão de uma fusão cps::lacZ.The spontaneous nodulation phenotype conferred by the snf2 allele is shown to be due to a cytokine-independent function of the mutant cytokine receptor LHK1 encoded by this allele, which is characterized by an L / F266 substitution in the CHASE domain (Figure 4). Inventive activity of wild-type and mutant Lotus LHK1 cytokine receptors and their response to cytokine was demonstrated by a two-component phosphor relocation assay developed in E. coli (Suzuki et al, (2001) Plant Cell Physio 42 : 107). In this assay cytokine induction can be determined without interference from endogenous cytokine levels, cytokine penetration, and interfering metabolic pathways present in plants. Phosphore relocation explored in this assay is usually involved in the regulation of bacterial extracellular polysaccharide synthesis by activating the operon cps in response to a sensor (RcsC) 1 which has a domain structure similar to plant cytokine receptors. Functional expression of a cytokine receptor in a deleted E. coli desensor RcsC strain allows cytokine uptake to be read as β-galactosidase activity by the expression of a cps :: lacZ fusion.

Consequentemente, os DNAcs Lhkl correspondentes ao alelode tipo selvagem e snf2, foram clonados no vetor de expressão plN-111 etransformados na cepa de sensor negativo SRC122 (ARcsC) abrigando ogene de cps::lacZ. Os transformantes foram cultivados durante a noite emLuria-ágar-ágar líquido e então colocados em placas LB1 suplementadascom e sem t-zeatina, quinetina, BAP (200 μΜ) cada um e tidiazuron (50 μΜ).Os meios LB foram ainda suplementados com X-Gal (5-bromo-4-cloro-3-indolil-p-D-galactosidase), um substrato para β-galactosidase. Após 40 horasde incubação à temperatura ambiente (25°C), as placas foram fotografadas.A cepa de E. coli (SRC122) foi tomada como um controle negativo. A ex-pressão do alelo snf2 (receptor de citocinina LHK1 mutante) é vista por indu-zir a atividade de β-galactosidase na ausência de citocinina (figura 5A). Aocontrário, a LHK1 de tipo selvagem induziu a atividade de β-galactosidase deuma maneira dependente de citocinina (figura 5A). O nível de atividade de β-galactosidase induzida pela expressão dos alelos de tipo selvagem e snf2 foiquantitativamente determinado nos transformantes depois do crescimentodurante a noite a 37°C em meio LB de acordo com o protocolo seguinte. Asculturas durante a noite foram diluídas 1:1000 em 50 ml de LB e, além disso,cultivadas durante 24 horas a 37°C. Depois do período de incubação, 1 mlda cultura foi centrifugado por 10 minutos a 13000 rpm. A DOeoo da culturarestante foi medida. O pélete foi ressuspenso em 1 ml de NaCl 0,85 %. 200μl da suspensão foram ajustados para 1 ml com o tampão Z (Guarentee(1983) Methods Enzymol., 101: 183). Duas gotas de SDS 0,1 % e 1 gota declorofórmio foram adicionadas. As amostras foram agitadas em vórtex e co-locadas em repouso a 30°C por 5 minutos. 200 μΙ de o-Nitrofenil-p-D-galactopiranosídeo ONPG (4 mg/ml) foram então adicionados à amostra,que foi ainda incubada a 30°C. Quando as amostras obtiverem uma cor a-marela apropriada (correspondente a DO420 = 0,300-0,600), a reação é para-da pela adição de 500 μl de Na2CO31 Μ. O tempo de parada foi registrado. ADO420 foi medida dentro de 1 hora por espectrofotometria. A atividade de β-galactosidase foi calculada como se segue: Unidades Relativas =(D0420*1000) / (D06oo*min*ml) (Guarentee, (1983) acima).Consequently, wild-type allelode and snf2 cDNAs Lhk1 were cloned into the pN-111 expression vector and transformed into the SRC122 (ARcsC) negative sensor strain harboring the cps :: lacZ gene. Transformants were cultured overnight on liquid Lauria-agar and then placed in LB1 plates supplemented with and without t-zeatin, quinetin, BAP (200 μΜ) each and tidiazuron (50 μΜ). LB media were further supplemented with X -Gal (5-bromo-4-chloro-3-indolyl-pD-galactosidase), a substrate for β-galactosidase. After 40 hours of incubation at room temperature (25 ° C), the plates were photographed. The E. coli strain (SRC122) was taken as a negative control. Expression of the snf2 allele (mutant cytokine receptor LHK1) is seen to induce β-galactosidase activity in the absence of cytokine (Figure 5A). In contrast, wild-type LHK1 induced β-galactosidase activity in a cytokine-dependent manner (Figure 5A). The level of β-galactosidase activity induced by expression of wild-type and snf2 alleles was quantitatively determined in transformants after growth overnight at 37 ° C in LB medium according to the following protocol. Overnight cultures were diluted 1: 1000 in 50 ml LB and further grown for 24 hours at 37 ° C. After the incubation period, 1 ml of the culture was centrifuged for 10 minutes at 13,000 rpm. The DOeoo of the culturestante was measured. The pellet was resuspended in 1 ml 0.85% NaCl. 200μl of the suspension was adjusted to 1 ml with Z buffer (Guarentee (1983) Methods Enzymol., 101: 183). Two drops of 0.1% SDS and 1 drop of chloroform were added. The samples were vortexed and placed at rest at 30 ° C for 5 minutes. 200 μΙ o-Nitrophenyl-p-D-galactopyranoside ONPG (4 mg / ml) was then added to the sample, which was further incubated at 30 ° C. When the samples obtain an appropriate a-marel color (corresponding to OD420 = 0.300-0.600), the reaction is stopped by the addition of 500 μl of Na2CO31 Μ. Stop time has been recorded. ADO420 was measured within 1 hour by spectrophotometry. Β-galactosidase activity was calculated as follows: Relative Units = (D0420 * 1000) / (D06oo * min * ml) (Guarentee, (1983) above).

Como mostrado na figura 5B, a expressão do alelo snf2 deLHK1 em células de E. coli induziu um nível três vezes maior da atividade deβ-galactosidase do que o controle, células de E. coli não transformadas ecélulas que expressam a LHK1 de tipo selvagem. A adição de citocinina re-sulta em uma indução de duas vezes da atividade de β-galactosidase emcélulas que expressam LHK1 enquanto células que expressam o alelo snf2respondem apenas com um aumento marginal na atividade. Esses resulta-dos demonstram que LHK1 é um receptor de citocinina, enquanto o receptormutante codificado pelo alelo snf2 é constitutivamente ativo, cuja atividade écomparável com a atividade induzida pela citocinina do receptor selvagem.Embora não pretenda ser vinculado pela teoria, é proposto que o domínioextracelular CHASE, que normalmente liga a citocinina para ativar a quinase(Kakimoto, (2001) Plant Cell Physiol. 42: 677; Anantharaman e Aravind,(2001) Trends Biochem Sei. 26: 579; Pas et al., (2004), FEBS Lett. 576: 287)é trancado em uma conformação ativa no receptor mutante (snf2). Essaspropriedades da LHK1 mutante explicariam tanto natureza genética domi-nante do alelo snf2 como os resultados do ensaio de fosforrealocação.As shown in Figure 5B, expression of the LHK1 snf2 allele in E. coli cells induced a three-fold higher level of β-galactosidase activity than control, untransformed E. coli cells and wild-type LHK1-expressing cells. The addition of cytokine results in a two-fold induction of β-galactosidase activity in LHK1-expressing cells while cells expressing the snf2 allele respond only with a marginal increase in activity. These results demonstrate that LHK1 is a cytokine receptor, while the mutant receptor encoded by the snf2 allele is constitutively active, whose activity is comparable to wild-type cytokine-induced activity. Although not intended to be bound by theory, it is proposed that the extracellular domain CHASE, which normally binds cytokine to activate kinase (Kakimoto, (2001) Plant Cell Physiol. 42: 677; Anantharaman and Aravind, (2001) Trends Biochem Sci. 26: 579; Pas et al., (2004), FEBS Lett. 576: 287) is locked into an active conformation at the mutant receptor (snf2). These properties of the mutant LHK1 would explain both the dominant genetic nature of the snf2 allele and the results of the phosphorealocation assay.

Exemplo 5. Impacto do alelo snf2 na regulação do crescimento e da diferen-ciação celularExample 5. Impact of the snf2 allele on cell growth and differentiation regulation

A conversão espontânea das células corticais da raiz em nódulosradiculares iniciais vista em mutantes snf2 é provavelmente devido à atividadeconstitutiva em uma ou mais etapas da via que controla a diferenciação celu-lar. A realização de cultura de tecido in vitro de células snf2 provê um meio deavaliar o impacto global do alelo snf2 em processos de diferenciação celulare/ou interferência com respostas normais a fito-hormônios. Assim sendo, hi-pocótilo e explantes de raiz (Figura 1 e 6) de 10 dias de idade de plantas detipo selvagem e snf2 (cultivados em >2 de meio salino B5) foram cultivados em36 proporções de ácido naftaleno acético (NAA) e 6-benzilaminopurina (BAP)durante três semanas. A faixa de concentrações de citocinina e auxina em-pregadas inclui níveis normalmente usados para a mais eficiente cultura invitro de explantes de tipo selvagem. Os explantes foram movidos para ummeio novo toda semana. Explantes representativos de cada combinação hor-monal foram arranjados em uma placa de Petri e fotografados.The spontaneous conversion of cortical root cells into initial root nodules seen in snf2 mutants is probably due to constitutive activity in one or more steps of the pathway that controls cell differentiation. In vitro tissue culture of snf2 cells provides a means of assessing the overall impact of the snf2 allele on processes of cell differentiation and / or interference with normal phytohormonal responses. Therefore, 10-day-old hypocotyl and root explants (Figures 1 and 6) of wild type and snf2 plants (grown in> 2 of B5 saline) were grown in 36 proportions of naphthalene acetic acid (NAA) and 6 -benzylaminopurine (BAP) for three weeks. The range of concentrations of cytokine and auxin employed includes levels commonly used for the most efficient invitro culture of wild-type explants. The explants were moved to a new medium every week. Representative explants of each hormonal combination were arranged in a petri dish and photographed.

Como mostrado na figura 1G, H e na figura 6, há pouca dife-rença observável no crescimento do calo em hipocótilo ou em explantes deraiz de plantas com alelos de tipo selvagem e snf2. Geralmente, a resposta àdose hormonal é semelhante. Somente em altas concentrações de auxina (2pg/ml NAA), explantes snf2 sobreviveram ligeiramente melhor do que explan-tes de tipo selvagem. De modo inverso, em acréscimo de citocinina (BAP) naausência de auxina, explantes snf2 desenvolveram menos calo do que os detipo selvagem. Quando cultivado em placas de cultura de tecido sem qualquerfito-hormônio, o calo verde cresceu a partir do final dos segmentos basais daraiz snf2, de uma maneira similar a uma resposta de tecido induzida pelacitocinina, e claramente diferente de segmentos de raiz de tipo selvagem(figura 11 e J). A cossegregação deste fenótipo do calo verde e do alelo snf2mostram que essa formação não induzida de calo radicular foi causada peloganho de função do receptor de citocinina codificado pelo alelo snf2.As shown in Fig. 1G, H, and Fig. 6, there is little observable difference in callus growth in hypocotyl or in deplorant explants of wild-type and snf2 alleles. Generally, the response to hormonal dose is similar. Only at high auxin concentrations (2pg / ml NAA) did snf2 explants survive slightly better than wild type explants. Conversely, in addition to cytokine (BAP) in the absence of auxin, snf2 explants developed less callus than wild type detectors. When cultured in tissue culture-free hormone-free plaques, green callus grew from the end of the snf2 root basal segments, in a manner similar to a phakocytokine-induced tissue response, and clearly different from wild-type root segments ( Figure 11 and J). The co-segregation of this green callus and snf2 allele phenotype shows that this uninduced root callus formation was caused by the cytokine receptor function encoded by the snf2 allele.

A microscopia em secções finas de raízes de tipo selvagem esnf2 (figura1 C,D e E,F) mostra pelo menos uma camada extra de célula devasos do xilema de raízes snf2. Depois de 6 dias da incubação em meio semhormônio, camadas adicionais de célula do periciclo originadas de divisõespericlinais foram observadas em conjunto com um aumento no número decélulas da vasculatura da raiz snf2 enquanto nenhuma modificação foi ob-servada em raízes de tipo selvagem (figura 1E, F).Thin section microscopy of wild type esnf2 roots (Figure 1 C, D and E, F) shows at least one extra layer of devas cell from snf2 root xylem. After 6 days of incubation in semhormonium medium, additional pericycle cell layers from periclinal divisions were observed together with an increase in the number of snf2 root vasculature cells while no changes were observed in wild type roots (Figure 1E, F)

Exemplo 6. Expressão do gene Lhkl em órgãos e dos genes Lhk1, Lrrb eNin em plantas Lotus de tipo selvagem e snf2 em resposta à citocininaExample 6. Expression of Lhkl gene in organs and Lhk1, Lrrb eNin genes in wild type and snf2 Lotus plants in response to cytokine

Os níveis no estado de equilíbrio dos transcritos Lhkl e ARR fo-ram determinados em raízes e outros órgãos da planta pela análise por RT-PCR quantitativa de acordo com o protocolo seguinte.Steady state levels of the Lhkl and ARR transcripts were determined in roots and other plant organs by quantitative RT-PCR analysis according to the following protocol.

Sementes de tipo selvagem e snf2 foram esterilizadas superfici-almente e germinadas como descrito anteriormente (Handberg, e Stougaard1(1992) Plant J. 2: 487). As plantas foram cultivadas por um período de 16hno claro/8h no escuro a 21 °C. O RNA total foi isolado de raízes, nódulos,folhas, flores e vagens de L. japonicus de tipo selvagem inoculado. O RNAmfoi extraído de raízes de plantas de tipo selvagem e snf2 cultivadas durante2 semanas em V* de B&D (Broughton e Dilworth, (1971) Biochem. J. 125:1075) suplementado com 0,5 μΜ de nitrato de potássio. As raízes foram tra-tadas com 10 μΜ de BAP em três pontos de tempo, 30 minutos, 3 horas e 8horas. As raízes não-tratadas foram tomadas como controle. RNAm tambémfoi extraído de segmentos de raiz de tipo selvagem e plantas snf2 cultivadasdurante 10 dias em meio salino B5 sem hormônio e meio salino B5 suple-mentado com 0,5 pg/ml de BAP. O kit Dynabeads mRNA direct (Invitrogen)foi usado para a extração de RNAm.Wild type and snf2 seeds were surface sterilized and germinated as previously described (Handberg, and Stougaard1 (1992) Plant J. 2: 487). The plants were grown for a period of 16h light / 8h in the dark at 21 ° C. Total RNA was isolated from roots, nodules, leaves, flowers and pods of inoculated wild type L. japonicus. MRNA was extracted from roots of wild-type and snf2 plants grown for 2 weeks in B&D V * (Broughton and Dilworth, (1971) Biochem. J. 125: 1075) supplemented with 0.5 μΜ potassium nitrate. Roots were treated with 10 μΜ BAP at three time points, 30 minutes, 3 hours and 8 hours. Untreated roots were taken as control. MRNA was also extracted from wild-type root segments and snf2 plants grown for 10 days in hormone-free B5 saline and B5 supplemented with 0.5 pg / ml BAP. The Dynabeads mRNA direct (Invitrogen) kit was used for mRNA extraction.

DNAc fita simples foi preparado usando transcriptase reversa(Fermentas). PCR quantitativa foi realizada em um Liqht Cvcler (Roche Mo-lecular Biochemicals) usando um Fast Start DNA master SYBR qreen kit(Roche) para amplificar os transcritos alvo a partir de 5 μΙ de DNAc diluído.Quatro genes constitutivos (Czechowski et aí., (2005) Genoma analysis 139:5) foram usados para determinar a expressão relativa dos genes alvo (vejaabaixo). Para cada tratamento, as razões relativas normalizadas dos genesalvo e os quatro genes constitutivos independentes foram calculadas usandoo Software de Quantificação Relativa da Roche (Quant). A média geométrica (Vandesompele et ai, (2002) Genome Biology 3:1) das razões da expressãorelativa para as três replicatas técnicas e biológicas e os intervalos de confi-ança correspondentes a 95% foram calculados como descrito anteriormente(Radutoiu et ai, (2003) Nature 425: 585).Single strand cDNA was prepared using reverse transcriptase (Fermentas). Quantitative PCR was performed on a Liqht Cvcler (Roche Molecular Biochemicals) using a Fast Start DNA master SYBR qreen kit (Roche) to amplify target transcripts from 5 μΙ of diluted cDNA. Four constitutive genes (Czechowski et al., (2005) Genome analysis 139: 5) were used to determine the relative expression of target genes (see below). For each treatment, the normalized relative ratios of the target genes and the four independent constitutive genes were calculated using Roche Relative Quantification Software (Quant). The geometric mean (Vandesompele et al, (2002) Genome Biology 3: 1) of the relative expression ratios for the three technical and biological replicates and the 95% confidence intervals were calculated as described above (Radutoiu et al. ( 2003) Nature 425: 585).

Os seguintes pares de iniciadores foram usados para a PCRThe following primer pairs were used for PCR

quantitativa:quantitative:

Seqüências dos iniciadores do gene constitutivo independente:Primers sequences of the independent constitutive gene:

1. PP24-TC9878-homólogo de AT1G13320-proteína fosfata-1. PP24-TC9878-AT1G13320-protein phosphate homologue

se 2Aif 2A

5'-GTAAATGCGTCTAAAGATAGGGTCC-3' 5'-ACTAGACTGTAGTGCTTGAGAGGC-3'5'-GTAAATGCGTCTAAAGATAGGGTCC-3 '5'-ACTAGACTGTAGTGCTTGAGAGGC-3'

2. UBC-CB828248 homólogo de AT5G25760- enzima conju-gadora de ubiquitina2. UBC-CB828248 homologue of AT5G25760- ubiquitin conjugating enzyme

5'-ATGTG CATTTTAAGAC AGGG-3'5'-GAACGTAGAAGATTGCCTGAA-3' 3. TB2C-BI418560- cadeia beta da tubulina5'-ATGTG CATTTTAAGAC AGGG-3'5'-GAACGTAGAAGATTGCCTGAA-3 '3. TB2C-BI418560- Tubulin beta chain

5'-GCTCACCACCCCAAGCTTTGG-3'5'-TGTCAATGGAGCAAACCCAACC-3'4. ATP-AW719841-ATP sintase-5'-GCTCACCACCCCAAGCTTTGG-3'5'-TGTCAATGGAGCAAACCCAACC-3'4. ATP-AW719841-ATP Synthase-

Si-Aacaccactctcgatcatttctctg-S'Si-Aacaccactctcgatcatttctctg-S '

5'-CAATGTCGCCAAGGCCCATGGTG-3'5'-CAATGTCGCCAAGGCCCATGGTG-3 '

Seqüências de iniciadores do gene alvo:LhklTarget gene primer sequences: Lhkl

5'-AATTTGGTGAACCGAAGGGTCGCCG-3'5'-TCGACGAGTGGCCTCAAACCCATCC-3' a/e/o snf2 de Lhkl5'-AATTTGGTGAACCGAAGGGTCGCCG-3'5'-TCGACGAGTGGCCTCAAACCCATCC-3 'to / and / Lhkl snf2

Snf2LC3 Fwd: 5'-AGAGGTCTTAAAGCCATTGTG-3'Snf2LC6Rev: 5-TATCAGGCTGAAATAATGCCG-3'Nin-AJ239041.Snf2LC3 Fwd: 5'-AGAGGTCTTAAAGCCATTGTG-3'Snf2LC6Rev: 5-TATCAGGCTGAAATAATGCCG-3'Nin-AJ239041.

5'-AGGAGCCCAAGTGAGTGCTA-3'5'-AGGAGCCCAAGTGAGTGCTA-3 '

5-GCCATCAAGGTATATGACGAG-3'5-GCCATCAAGGTATATGACGAG-3 '

ARR5-CB827384-homólogo de AT3G48100-ARR5ARR5-CB827384-AT3G48100-ARR5 homologue

5'-TCTTGACTCGAATTGATAGGTGC-3'5'-TCTTGACTCGAATTGATAGGTGC-3 '

5'-GATAGAGATGGCCTGCAACTACTG-3'5'-GATAGAGATGGCCTGCAACTACTG-3 '

A análise do transcrito mostra Lhkl a ser expresso em níveismais altos em raízes, nódulos e folhas, mas os transcritos estiveram presen-tes em todos os órgãos testados (figura 7A).Transcript analysis shows Lhkl being expressed at higher levels in roots, nodules and leaves, but transcripts were present in all organs tested (Figure 7A).

Modificações induzidas pela citocinina em processos celularesem plantas são acompanhadas pela expressão aumentada de genes regula-dores de resposta (ARR) que pertencem à classe Tipo-A (Hutchison e Kie-ber, (2002) The plant Cell 14: 47). Dez genes que codificam ARRs em Arabi-dopsis e os seus transcritos foram encontrados em todos os tecidos adultos.Os genes ARR Tipo-A são transcricionalmente induzidos pela citocinina eARR4 e ARR5 são genes de resposta primária rapidamente induzidos. Umavez que a proteína receptora mutante codificada pelo alelo snf2 mostrou ati-vidade constitutiva no sistema de teste em E. coli, a ativação de um LotusARR5 homólogo (denominado Lrr5) foi determinado em raízes snf2 e de tiposelvagem (figura 7). Um nível duas vezes maior do transcrito Lrr5 foi encon-trado em explantes de raiz de mutantes snf2 incubados no meio B5 semhormônio em comparação com explantes de tipo selvagem, enquanto a adi-ção de citocinina [BAP] ao meio B5 aumentou duas a três vezes o nível detranscrito Lrr5 em ambos os explantes, snf2 e de tipo selvagem (figura 7B).O tratamento direto das raízes com citocinina também aumentou o nível detranscrito Lrr5 em raízes snf2 e de tipo selvagem, mas nesse experimento,uma diferença na expressão entre raízes snf2 não-tratadas e de tipo selva-gem não foi encontrada (figura 7C).Cytokine-induced modifications in plant cellular processes are accompanied by increased expression of Type-A regulatory response genes (ARRs) (Hutchison and Kie-ber, (2002) The plant Cell 14: 47). Ten genes encoding Arabi-dopsis ARRs and their transcripts have been found in all adult tissues. Type-A ARR genes are transcriptionally induced by eARR4 and ARR5 cytokines are rapidly induced primary response genes. Since the mutant receptor protein encoded by the snf2 allele showed constitutive activity in the E. coli test system, activation of a homologous LotusARR5 (named Lrr5) was determined in snf2 and wild type roots (Figure 7). A two-fold higher level of the Lrr5 transcript was found in root explants of snf2 mutants incubated in semhormone B5 compared to wild-type explants, while cytokine [BAP] addition in B5 medium increased two to three times. Lrr5 transcript level in both snf2 and wild-type explants (Figure 7B). Direct treatment of cytokine roots also increased Lrr5 transcript level in snf2 and wild-type roots, but in this experiment, a difference in expression between roots untreated and jungle-type snf2 was not found (figure 7C).

Os transcritos do gene Lhkl também são vistos aumentandorapidamente em resposta ao tratamento com citocinina no tipo selvagem emutantes snf2 (figura 7D), que é compatível com a inducibilidade da citocini-na de seu homólogo em Arabidopsis (gene AHK4). O gene Nin em Lotus,que é necessário para a iniciação do nódulo primordial (Figura 5C), é tam-bém transcricionalmente super-regulado pela citocinina (figura 7E), mas osníveis do transcrito em raízes snf2 não-tratadas e tratadas não foram signifi-cativamente diferentes daqueles de raízes de tipo selvagem. A expressãoectópica de Nin em explantes mutantes de raiz snf2 incubados em meio B5sem hormônio não foi detectada (Fig. 2F).The Lhkl gene transcripts are also rapidly increasing in response to snf2-emutant wild-type cytokine treatment (Figure 7D), which is compatible with its homologous cytokine inducibility in Arabidopsis (AHK4 gene). The Nin gene in Lotus, which is required for primordial nodule initiation (Figure 5C), is also transcriptionally over-regulated by cytokine (Figure 7E), but transcript levels in untreated and treated snf2 roots were not significant. -ctively different from those of wild type roots. Nin ectopic expression in snf2 mutant root explants incubated in hormone-free B5 medium was not detected (Fig. 2F).

Em Arabidopsis, a via de resposta ARR é dessensibilizada de-pois da exposição prolongada à citocinina (Rashotte et al., (2003), PlantPhysiol. 132: 1998), que pode explicar a relativamente pequena ou a falta desuper-regulação de Lrr5 e Nin em raízes de mutante snf2.In Arabidopsis, the ARR response pathway is desensitized after prolonged cytokine exposure (Rashotte et al. (2003), PlantPhysiol. 132: 1998), which may explain the relatively small or lack of regulation of Lrr5 and Nin in snf2 mutant roots.

Exemplo 7. O hábito de crescimento da planta Lotus Snf2 é sensível à cito-cininaExample 7. The growth habit of the Lotus Snf2 plant is cytokine sensitive

Embora as modificações transcricionais na planta Lotus snf2 se-jam limitadas, o crescimento da planta foi visto ser fortemente afetado pelaaplicação de citocinina fornecida externamente. Assim sendo, as plantas detipo selvagem e snf2 foram cultivadas em 1/2 de salina B5 com e sem aumen-to das concentrações de BAP durante 3 semanas. O comprimento do broto eda raiz de pelo menos 60 plantas de cada tratamento foram medidos. Deacordo com a atividade constitutiva do receptor de citocinina mutante codifi-cado pelo alelo snf2, o crescimento do broto e da raiz da planta snf2 foramhipersensíveis à citocinina em comparação ao de tipo selvagem (figura 8).Exemplo 8.O fenótipo de raiz snf2 cosseqreqado com a mutação snf2Although transcriptional modifications in the Lotus snf2 plant are limited, plant growth has been seen to be strongly affected by externally supplied cytokine application. Therefore, wild type and snf2 plants were grown in 1/2 of B5 saline with and without increasing BAP concentrations for 3 weeks. Shoot length and root length of at least 60 plants from each treatment were measured. According to the constitutive activity of the snf2 allele-encoded mutant cytokine receptor, the shoot and root growth of the snf2 plant were hypersensitive to wild-type cytokine (Figure 8). Example 8. The snf2 root phenotype followed with snf2 mutation

As raízes da plantas snf2, cultivada em 1/2 de meio salino B5sem hormônio, são caracterizados pelo aumento da divisão celular no perici-cio e tecido vascular (figura 1 D,F). Este padrão de crescimento, que leva aum fenótipo de raiz inchada, é atribuído à expressão do alelo snf2. Para con-firmar que o fenótipo de raiz inchada de fato co-segrega com o fenótipo denodulação espontânea, o alelo mutante snf2-2 foi retrocruzada com o ecoti-po Gifu de tipo selvagem e as plantas F2 [50] foram cultivadas em !4 demeio B&D durante tempo suficiente para permitir o registro do fenótipo denodulação espontânea (~5 semanas). As raízes das plantas noduladas enão-noduladas foram cultivadas durante 3 semanas em meio sem hormônio,e então registradas para a presença e a ausência de inchaço radicular. Asplantas de tipo selvagem e snf2 foram tomadas como controles. Somente asraízes das plantas F2 que desenvolveram nódulos espontâneos mostraram oinchaço dos segmentos radiculares que foi comparável ao inchaço dos seg-mentos da raiz snf2. Isto indica que a expressão da proteína Lhkl indepen-dente de citocinina mutante nas plantas Lotus confere à raiz e ao broto snf2o fenótipo de crescimento e o fenótipo de nodulação espontânea.The roots of snf2 plants, grown in 1/2 of B5-free saline medium without hormone, are characterized by increased cell division in the pericardium and vascular tissue (Figure 1 D, F). This growth pattern, which leads to a swollen root phenotype, is attributed to the expression of the snf2 allele. To confirm that the swollen root phenotype actually co-secretes with the spontaneous denodulation phenotype, the snf2-2 mutant allele was back-crossed with the wild-type Gifu ecotype and F2 plants [50] were grown in! 4 B&D for sufficient time to allow spontaneous deodulation phenotype to be recorded (~ 5 weeks). The roots of the nodulated and non-nodulated plants were grown for 3 weeks in hormone-free medium, and then recorded for the presence and absence of root swelling. Wild type and snf2 plants were taken as controls. Only roots of F2 plants that developed spontaneous nodules showed swelling of root segments that was comparable to swelling of snf2 root segments. This indicates that expression of mutant cytokine-independent Lhkl protein in Lotus plants confers to the root and bud snf2 the growth phenotype and the spontaneous nodulation phenotype.

Exemplo 9. snf2 e a sinalização de citocinina atuam a jusante na via detransdução de sinal induzida pelo do fator Nod.Example 9. snf2 and cytokine signaling act downstream in the Nod factor-induced signal transduction pathway.

O fenótipo dos mutantes snf2 sugere que a sinalização de cito-cinina seja parte de, ou atue a jusante, da via de transdução de sinal induzi-da pelo fator Nod (figura 5C). Para testar esta hipótese, o construto do genesnf2 foi transformado nas plantas mutantes que transportam mutações emgenes que constituem a via comum de transdução de sinal compartilhadacom fungos micorrizais ou nas plantas mutantes defeituosas em genes amontante ou a jusante.The snf2 mutant phenotype suggests that cytokine signaling is part of, or downstream of, the Nod factor-induced signal transduction pathway (Figure 5C). To test this hypothesis, the nf2 gene construct was transformed into mutant plants carrying gene mutations that constitute the common pathway of shared signal transduction with mycorrhizal fungi or defective mutant plants into bulging or downstream genes.

Assim sendo, sete L. japonicus único e duplo mutantes de nodu-lação foram transformados usando Agrobacterium rhizogenes que transpor-tam o construto mutante do gene snf2 ou um vetor vazio como descrito ante-riormente (Stougaard, 1995 acima). Todos os mutantes exceto sy/r?35 foramgenotipados por sequenciamento dos produtos de PCR que cobrem a muta-ção usando os seguintes iniciadores:Thus seven seven single and double nodulation mutants were transformed using Agrobacterium rhizogenes which transpose the snf2 gene mutant construct or an empty vector as described above (Stougaard, 1995 above). All mutants except sy / r? 35 were genotyped by sequencing the mutation-covering PCR products using the following primers:

nfr1nfr1

Nfr1-1 Msel CAPSfw(56) 5'-Nfr1-1 Msel CAPSfw (56) 5'-

CGCTGGTTTACCCATAAACGTGTTC-3'CGCTGGTTTACCCATAAACGTGTTC-3 '

Nfr1-1 Msel CApsrv(SS)Sl-GGGCAAATGCATTTGTGCTGAG-S'nfr5Nfr1-1 Msel CApsrv (SS) Sl-GGGCAAATGCATTTGTGCTGAG-S'nfr5

K2fwB 5'-CCAGCTAGGTGATAGCTACG-3'K2revC 5'-CCAGAAGATGAATGCTGCTTT-3'K2fwB 5'-CCAGCTAGGTGATAGCTACG-3'K2revC 5'-CCAGAAGATGAATGCTGCTTT-3 '

S5R rev5 (64) 5'-GGTATTAGAACGCCCCCTGG-3'S5R rev5 (64) 5'-GGTATTAGAACGCCCCCTGG-3 '

symRK (tipo selvagem)symRK fw1 5'-CTGAGTTTGGACCCCTTTTG-3'symRK revi 5'-ACGCCCTTATGAAAATGTGG-3'symRK (gene mutante)symRK (wild type) symRK fw1 5'-CTGAGTTTGGACCCCTTTTG-3'symRK revi 5'-ACGCCCTTATGAAAATGTGG-3'symRK (mutant gene)

Lore2 5' LTR fora P 5-GGAGCTCTGATACCAATGTTAGG-3'cac41.5F 5'-CGGCAATAGAGCGCTGGAGAGTTG-3'ccamkLore2 5 'LTR Out P 5-GGAGCTCTGATACCAATGTTAGG-3'cac41.5F 5'-CGGCAATAGAGCGCTGGAGAGTTG-3'ccamk

LTsymI5 (60) fw 5-TATGACACAGATAGATCAGGG-3'LTsymI5 (60) rev 5'-GAGAGCGGCTCAATGAATGT-3'ninLTsymI5 (60) fw 5-TATGACACAGATAGATCAGGG-3'LTsymI5 (60) rev 5'-GAGAGCGGCTCAATGAATGT-3'nin

Resc 5'-CTCAGAGCACGCTTCTTGGA-3 futuros'Resc rev 5'-ATCATGTGTGCAATCCATGATG-3'har1Resc 5'-CTCAGAGCACGCTTCTTGGA-3 Futures'Resc rev 5'-ATCATGTGTGCAATCCATGATG-3'har1

#3fw2 - 5'-CCTGAAATGCCTATTCGTTGAG-3'#3rev2 5'-CACAGCTTCTTCTGCATGCG-3'snfl# 3fw2 - 5'-CCTGAAATGCCTATTCGTTGAG-3 '# 3rev2 5'-CACAGCTTCTTCTGCATGCG-3'snfl

Ca2fwd Si-Tggcttgcatccaaacggc-S'Ca2fwd Si-Tggcttgcatccaaacggc-S '

Ca3-3rev 5-ACTATTGTTGTCTCACTTTAGTG-3'snf2Ca3-3rev 5-ACTATTGTTGTCTCACTTTAGTG-3'snf2

C3fwd 5'-TGGGATAATTGGTTGCTTGACA-3'C3Brev 5-TGACAATGTGAGTTCCAGCAG-3'A raízes cabeludas transgênicas foram então monitoradas paranodulação espontânea na ausência do rizóbio.C3fwd 5'-TGGGATAATTGGTTGCTTGACA-3'C3Brev 5-TGACAATGTGAGTTCCAGCAG-3'A transgenic hairy roots were then monitored for spontaneous nodulation in the absence of rhizobia.

Tabela 3Table 3

Nodulação espontânea em plantas Lotus não-nodulada mutantesSpontaneous nodulation in mutant non-nodulated Lotus plants

transformadas com o gene snf2.transformed with the snf2 gene.

<table>table see original document page 30</column></row><table>As raízes cabeludas de mutantes transformados foram registradas na au-sência da Μ. Ioti. As raízes das plantas mutantes controle transformadascom um vetor vazio não desenvolveram nódulos espontâneos.<table> table see original document page 30 </column> </row> <table> The hairy roots of transformed mutants were recorded in the absence of Μ. Ioti The roots of mutant control plants transformed with an empty vector did not develop spontaneous nodules.

A nodulação espontânea observada em nfr1-1, nfr5-2 e nfr1-1nfr5-2 receptor de fator Nod, único e duplo mutantes perdendo a reposta ele-trofisiológica precoce ao fator Nod (Radutoiu et ai, (2003) acima; Madsen etai, (2003) acima), demonstra que Lhkl funciona a jusante da captação desinalização do fator Nod. As vias comuns mutantes symRK desprovidas derápida oscilação citoplasmática de Ca2+ (Stracke et ai, (2002) Nature, 417:959; Niwa et ai, (2001) Mol Plant Microbe Internet. 14: 848) e os ccamk mu-tantes sugerem ser incapazes de interpretar a rápida oscilação citoplasmáti-ca de Ca2+ (Tirichine et ai, (2006) Nature 441: 1153), também desenvolvemnódulos radiculares espontâneos em raízes transgênicas transformadas como construto do gene snf2. Nos mutantes nin e sym35 que são suprimidosantes da iniciação da divisão celular induzida através da via comum, nenhumnódulo espontâneo foi observado em raízes transgênicas, mostrando que acaptação da sinalização de citocinina atua a montante do início da divisãocelular, ou funciona em uma via paralela (figura 5C). Evidências adicionaisde um papel central da citocinina e da captação de citocinina na via detransdução de sinal induzida pelo fator Nod amplificada através de uma viacomum vêm do efeito aditivo das mutações snf1-1 e snf2 em duplo mutan-tes. Os mutantes snf1-1 sintetizam uma proteína CCaMK defeituosa em au-tofosforilação (Tirichine et ai, (2006) acima) e desenvolvem uma média desete nódulos espontâneos. Mutantes snf2 desenvolvem uma média de trêsnódulos espontâneos, enquanto snf1-1 snf2 duplo mutantes excedem ambose desenvolvem aproximadamente dezessete nódulos espontâneos. A con-versão de células corticais em células tronco de nódulo ou o subsequentedesenvolvimento de órgão parecem, por isso, rigidamente controlados. Essateoria foi testada pelo cruzamento do alelo snf2 em um contexto genético dehipernodulação har1-1 (Krusell et ai, (2002) Nature 420: 422). Na ausênciado rizóbio, snf2 har1-1 duplo mutantes homozigotos desenvolveram umamédia de quatorze nódulos espontâneos, enquanto os mutantes snf2 desen-volveram uma média de três, e har1-1 nenhum (figura 9). Esses resultadosindicam que somente algumas poucas células se desdiferenciaram ou quesomente poucas células desdiferenciadas sustentam as divisões celularesdurante o processo de iniciação do nódulo snf2. A autorregulação controladapelo broto do número de nódulos radiculares (Krusell et al., (2002) Nature420: 422) está assim atuando a jusante da sinalização de citocinina e a cito-cinina induziu a ativação das células iniciadoras do nódulo radicular (figura5C). Os dados claramente demonstram que a sinalização de citocinina é ne-cessária e suficiente para a desdiferenciação e a proliferação celular levandoà formação de nódulo radicular e que este processo de desenvolvimentopode ser provocado independentemente da citocinina pela expressão doalelo snf2 dominante. Surpreendentemente, o fenótipo conferido pela ex-pressão do alelo snf2, expresso sob controle de um promotor regulado (porexemplo, promotor do gene Lhk1) é caracterizado pela organogênese con-trolada de nódulos radiculares, em vez de proliferação celular descontrolada,independente da citocinina.Spontaneous nodulation observed in nfr1-1, nfr5-2 and nfr1-1nfr5-2 Nod factor receptor, single and double mutants losing early electrophysiological response to Nod factor (Radutoiu et al, (2003) above; Madsen etai, (2003) above), demonstrates that Lhkl works downstream of the Nod factor uptake. Common symRK mutant pathways lacking rapid cytoplasmic oscillation of Ca2 + (Stracke et al., (2002) Nature, 417: 959; Niwa et al. (2001) Mol Plant Microbe Internet. 14: 848) and mutant ccamk suggest being incapable. to interpret the rapid cytoplasmic oscillation of Ca2 + (Tirichine et al. (2006) Nature 441: 1153), also developing spontaneous root modules in transformed transgenic roots as a snf2 gene construct. In the nin and sym35 mutants that are suppressed from the initiation of common-cell induced cell division, no spontaneous modulus has been observed in transgenic roots, showing that cytokine signaling uptake acts upstream of cell division onset or works in a parallel pathway (figure 5C). Further evidence of a central role of cytokine and cytokine uptake in the Nod factor-induced signal transduction pathway amplified via a common pathway comes from the additive effect of the double mutant snf1-1 and snf2 mutations. The snf1-1 mutants synthesize a defective auto-phosphorylation CCaMK protein (Tirichine et al., (2006) above) and develop an average of seven spontaneous nodules. Snf2 mutants develop an average of three spontaneous nodules, while snf1-1 snf2 double mutants exceed ambose and develop approximately seventeen spontaneous nodules. Conversion of cortical cells into nodule stem cells or subsequent organ development seems therefore tightly controlled. Theory has been tested by crossing the snf2 allele in a har1-1 hypermodulation genetic context (Krusell et al, (2002) Nature 420: 422). In the absence of rhizobia, snf2 har1-1 double homozygous mutants developed an average of fourteen spontaneous nodules, while snf2 mutants developed an average of three, and har1-1 none (Figure 9). These results indicate that only a few cells have been undifferentiated or that only a few undifferentiated cells support cell division during the snf2 nodule initiation process. Self-regulation controlled by the bud of the number of root nodules (Krusell et al. (2002) Nature420: 422) is thus acting downstream of cytokine signaling and cytokinin induced activation of the root nodule initiation cells (Figure 5C). The data clearly demonstrate that cytokine signaling is necessary and sufficient for cell de-differentiation and proliferation leading to root nodule formation and that this developmental process can be triggered independently of cytokine by expression of the dominant snf2 allele. Surprisingly, the phenotype conferred by snf2 allele expression, expressed under the control of a regulated promoter (eg, the Lhk1 gene promoter) is characterized by uncontrolled root-cell organogenesis rather than uncontrolled cytokine-independent cell proliferation.

Exemplo 10. Kit de detecção para distinguir o alelo mutante snf2 do alelo detipo selvagem que codifica a histidina quinase LKH1Example 10. Detection kit for distinguishing mutant snf2 allele from wild-type histidine kinase encoding allele LKH1

Um kit para a detecção do alelo mutante snf2 útil para o rastre-amento de genótipos alvo compreende pelo menos:A snf2 mutant allele detection kit useful for screening target genotypes comprises at least:

Dois grupos alternativos de dCAPS (seqüência polimórfica am-plificada clivada derivada) iniciadores são providos para a detecção do tiposelvagem versus a mutação:Two alternative groups of dCAPS (derived cleaved amplified polymorphic sequence) primers are provided for detection of wild type versus mutation:

Grupo um:Group one:

Snf2 Xmnl/Asp700 dCAPS fw(74)Snf2 Xmnl / Asp700 dCAPS fw (74)

5'-CACCAAAATTGCTTGGTTACCAGCAAGTTGACCGA5'-CACCAAAATTGCTTGGTTACCAGCAAGTTGACCGA

Snf2 Xmnl/Asp700 dCAPS rv (69) 5'-CCCTTCATGTGGCCCTTACCCAACSnf2 Xmnl / Asp700 dCAPS rv (69) 5'-CCCTTCATGTGGCCCTTACCCAAC

Os iniciadores são convenientes para o uso em um teste PCRrealizado com DNA genômico como molde, onde a amplificação é realizadaa 60°C com 30 segundos de alongamento e 35-40 ciclos. O produto de 225pb é clivado pela Xmnl/Asp700 no mutante, e não é clivado no tipo selva-gem. Os produtos de PCR clivados podem ser identificados depois de suaseparação em gel de agarose a 1 %.Primers are suitable for use in a PCR assay performed with genomic DNA as a template, where amplification is performed at 60 ° C with 30 seconds of elongation and 35-40 cycles. The 225bp product is cleaved by the Xmnl / Asp700 in the mutant, and is not cleaved in the jungle type. Cleaved PCR products can be identified after their separation on 1% agarose gel.

Grupo dois:Group two:

Snf2 Rsal dCAPS fw (72)Snf2 Rsal dCAPS fw (72)

5'-GATCCTTTGATGTTGAGTCCCTTGTGGAGAATGTA Snf2 Rsal dCAPS rv (62)5'-5'-GATCCTTTGGGTTGAGTCCCTTGTGGAGAATGTA Snf2 Rsal dCAPS rv (62) 5'-

CTGAAACTCAGAAAATGTACTACAACCTGAAACTCAGAAAATGTACTACAAC

Os iniciadores são convenientes para o uso em um teste PCRrealizada com DNA genômico como molde, onde a amplificação é realizadaa 48°C com 30 segundos de alongamento e 35-40 ciclos. O produto de pb éclivado por bu Rsal no tipo selvagem, mas não é clivado no mutante.Primers are convenient for use in a PCR assay performed with genomic DNA as a template, where amplification is performed at 48 ° C with 30 seconds of elongation and 35-40 cycles. The bp product is cleaved by bu Rsal in the wild type, but is not cleaved in the mutant.

Os produtos de PCR clivados podem ser identificados depois desua separação em gel de agarose a 1%.Cleaved PCR products can be identified after their separation on 1% agarose gel.

Exemplo 11. Iniciadores de amplificação para clonagem da seqüência decodificação do DNAc do snf2 .Example 11. Amplification primers for cloning the snf2 cDNA decoding sequence.

Os iniciadores para amplificar a seqüência de codificação dapartida para parada (DNAc) são:The primers for amplifying the stop-start coding sequence (cDNA) are:

Snf2 DNAc fw ATGGGTCTTGGGTTCAAGATGCASnf2 DNAc fw ATGGGTCTTGGGTTCAAGATGCA

Snf2 DNAc rev TCATGAGTCTGAAGCAGGCTTGGSnf2 cDNA rev TCATGAGTCTGAAGCAGGCTTGG

A PCR foi realizada com o anelamento a 58°C e tempo de alon-gamento de 3 minutos e 20 segundos por 25 ciclos com o clone de DNAccomo molde.LISTAGEM DE SEQÜÊNCIAPCR was performed with annealing at 58 ° C and stretching time of 3 minutes and 20 seconds for 25 cycles with the cDNA clone as template. SEQUENCE LISTING

<110> Aarhus universitet, LedelsessekretariatetTiri chine, Leila<110> Aarhus universitet, LedelsessekretariatetTiri chine, Leila

<120> MOLÉCULA DE DNA QUE CODIFICA UM POLIPEPTÍDEO DE HISTIDINA QUI-<120> DNA Molecule that encodes a HISTIDINE CHIP polypeptide

NASE MUTANTE E SEU USO RELACIONADO, POLIPEPTÍDEO DE HISTIDINAQUINASE MUTANTE, BEM COMO MÉTODO DE PRODUÇÃO DE UMA PLANTA GE-NETICAMENTE MODIFICADAMUTANT NASE AND ITS RELATED USE, MUTANT HISTIDINAKINASE POLYPEPTIDE, AS A METHOD OF PRODUCTION OF A GE-NETICALLY MODIFIED PLANT

<130> P80600597DK00<130> P80600597DK00

<160> 20<160> 20

<170> Patentln version 3.2<170> Patentln version 3.2

<210> 1<210> 1

<211> 12738<211> 12738

<212> DNA<212> DNA

<213> Lotus japonicus<213> Lotus japonicus

<220><220>

<221> promoter<221> promoter

<222> (1)..(2359)<222> (1) .. (2359)

<223> promoter + 5' UTR of transcript<220><223> promoter + 5 'UTR of transcript <220>

<221> exon<221> exon

<222> (2360)..(2704)<222> (2360) .. (2704)

<223> start of CDS in Ist exon<220><223> start of CDS in Ist exon <220>

<221> Intron<221> Intron

<222> (2705)..(3024)<222> (2705) .. (3024)

<220><220>

<221> exon<221> exon

<222> (3025)..(3258)<220><222> (3025) .. (3258) <220>

<221> Intron<221> Intron

<222> (3259)..(3390)<220><221> exon<222> (3259) .. (3390) <220> <221> exon

<222> (3391)..(3560)<222> (3391) .. (3560)

<220><220>

<221> Intron<221> Intron

<222> (3561)..(3834)<220><222> (3561) .. (3834) <220>

<221> exon<221> exon

<222> (3835)..(4053)<220><222> (3835) .. (4053) <220>

<221> Intron<221> Intron

<222> (4054)..(4145)<220><222> (4054) .. (4145) <220>

<221> exon<221> exon

<222> (4146)..(4332)<220><222> (4146) .. (4332) <220>

<221> Intron<221> Intron

<222> (4333)..(4420)<222> (4333) .. (4420)

<220><220>

<221> exon<221> exon

<222> (4421)..(4472)<220><222> (4421) .. (4472) <220>

<221> Intron<221> Intron

<222> (4473)..(4586)<220><222> (4473) .. (4586) <220>

<221> exon<221> exon

<222> (4587)..(4816)<220><222> (4587) .. (4816) <220>

<221> Intron<221> Intron

<222> (4817)..(5037) <220><222> (4817) .. (5037) <220>

<221> exon<221> exon

<222> (5038)..(5136)<220><222> (5038) .. (5136) <220>

<221> Intron<222> (5137)..(5308)<220><221> exon<221> Intron <222> (5137) .. (5308) <220> <221> exon

<222> (5309)..(6024)<220><222> (5309) .. (6024) <220>

<221> Intron<222> (6025)..(7042)<220><221> Intron <222> (6025) .. (7042) <220>

<221> exon<222> (7043)..(7513)<220><221> exon <222> (7043) .. (7513) <220>

<221> Intron<222> (7514)..(8306)<220><221> Intron <222> (7514) .. (8306) <220>

<221> exon<222> (8307)..(8564)<220><221> Intron<221> exon <222> (8307) .. (8564) <220> <221> Intron

<222> (8565)..(12738)<222> (8565) .. (12738)

<223> 3"UTR + 3 prime of transcript<223> 3 "UTR + 3 prime of transcript

<400> 1<400> 1

cccgggattg ttcatatccc catgtgtgtg tttggtttgg acacaagcaa gcttgccaca 60gtggtgttta caactcattc tgaattatat atatatatat atatatataa ctaatctcta 120tcaataattt agttagtttt atgggataat atgattcccg tcaaattagc ttgtaggtta 180aaaatccata tttatataat aaattagact tatctaaatc acttaagtaa atatgatgta 240aaatcacaat agcttaggta gtttaacaat gttttatcat gaatttaaaa aaaattagtt 300gtttcaaaat attattggac caaataagca aagaggagct aatccaatta taaacagcac 360atctttataa ttgttttcta ctcgatctaa tattggttcc ttctttcctt gagaaaaaaa 420gggttaaaaa tgcgatttgt tttgtaaaat taatttgtga cattaacaag agaaaactct 480acttttttaa tttttaagaa gttacttaaa tatagaaaga aagaagttac ttaaatatga 540aacttgtcta accaaagtca accgatcaaa gcgtttggtt agcaaagaga acttaaaaaa 600cccgggattg ttcatatccc catgtgtgtg tttggtttgg acacaagcaa gcttgccaca 60gtggtgttta caactcattc tgaattatat atatatatat atatatataa ctaatctcta 120tcaataattt agttagtttt atgggataat atgattcccg tcaaattagc ttgtaggtta 180aaaatccata tttatataat aaattagact tatctaaatc acttaagtaa atatgatgta 240aaatcacaat agcttaggta gtttaacaat gttttatcat gaatttaaaa aaaattagtt 300gtttcaaaat attattggac caaataagca aagaggagct aatccaatta taaacagcac 360atctttataa ttgttttcta ctcgatctaa tattggttcc ttctttcctt gagaaaaaaa 420gggttaaaaa tgcgatttgt tttgtaaaat taatttgtga cattaacaag agaaaactct 480acttttttaa tttttaagaa gttacttaaa tatagaaaga aagaagttac ttaaatatga 540aacttgtcta accaaagtca accgatcaaa gcgtttggtt agcaaagaga acttaaaaaa 600

ataactaaat taactgtttg aacagtgatg taacagagaa atagaatggc ttacatatgg 660ataactaaat taactgtttg aacagtgatg taacagagaa atagaatggc ttacatatgg 660

attatggagt taatggaagg cacactagtt ttagttcacg gcaacaagac agaattatcc 720attatggagt taatggaagg cacactagtt ttagttcacg gcaacaagac agaattatcc 720

tcctatattt taggaggttt tcttatttat ttttaatgta agacgtgtat ccttcttttg 780tcctatattt taggaggttt tcttatttat ttttaatgta agacgtgtat ccttcttttg 780

aggtaataat gttcgagctt gaaacattca cattatagtg tgactaattg cttaatatga 840aggtaataat gttcgagctt gaaacattca cattatagtg tgactaattg cttaatatga 840

atcaagcatg taccacttga aacactcact catcgtttat aatataatta ctttatagta 900atcaagcatg taccacttga aacactcact catcgtttat aatataatta ctttatagta 900

ctataaagta tgtaattatt ttctcctttt taagtatctc aatcattttt ttgacccaaa 960ctataaagta tgtaattatt ttctcctttt taagtatctc aatcattttt ttgacccaaa 960

tattatttta gagaagaaac atgtcttaag accaatacat caacatatat ggagagtagt 1020tattatttta gagaagaaac atgtcttaag accaatacat caacatatat ggagagtagt 1020

ttgacaacct ctcatgagct ttctaaagcc aaacaaatgt actcacataa aaaaattaaa 1080ttgacaacct ctcatgagct ttctaaagcc aaacaaatgt actcacataa aaaaattaaa 1080

aatttacagc aagtgagtta taaagagtga ttaataaatg ttgaataacg atggagggta 1140aatttacagc aagtgagtta taaagagtga ttaataaatg ttgaataacg atggagggta 1140

gttatgaaat tccataaata aagcaaagga tgtttatggc aattgacatg ggaataagat 1200gttatgaaat tccataaata aagcaaagga tgtttatggc aattgacatg ggaataagat 1200

ccgcgcgttg tcgccgtgtc ataatcgctc agatttgtga tagcgagaga tttccattct 1260ccgcgcgttg tcgccgtgtc ataatcgctc agatttgtga tagcgagaga tttccattct 1260

tttcctcttt tttcgaacga acgaacgaac gaacacagta gcagctgtat taggattcag 1320tttcctcttt tttcgaacga acgaacgaac gaacacagta gcagctgtat taggattcag 1320

attgcatgat acattgatat tgatattgat attgatattg atattgttgt gggtcaagtc 1380attgcatgat acattgatat tgatattgat attgatattg atattgttgt gggtcaagtc 1380

tctactctac tatccaaagt aagcatatat atagagagag agagagagct tgggactagg 1440tctactctac tatccaaagt aagcatatat atagagagag agagagagct tgggactagg 1440

gaggatatta gcttatgtga ctgtgaagtt gaagggagac aagagcgtat ctggcaaaat 1500gaggatatta gcttatgtga ctgtgaagtt gaagggagac aagagcgtat ctggcaaaat 1500

cctcaaaata aaatactagt actagtacag agaaaagaga ctaagagaga gagtgctgct 1560cctcaaaata aaatactagt actagtacag agaaaagaga ctaagagaga gagtgctgct 1560

gcacatcaag acccattgtg atttgtgatt tgtgatttgt gatttgtgat ttgtgagtga 1620gcacatcaag acccattgtg atttgtgatt tgtgatttgt gatttgtgat ttgtgagtga 1620

gttcattgta caggtattat tgtttgttgt ttctctcctc aaccaccctc taaagtctaa 1680gttcattgta caggtattat tgtttgttgt ttctctcctc aaccaccctc taaagtctaa 1680

tctaactcat tgggctctgt gcttagctgg ttgtgttttg tgtatggtga attagggggt 1740tctaactcat tgggctctgt gcttagctgg ttgtgttttg tgtatggtga attagggggt 1740

caatctctgg ttttcatcat tattatatta tatatgggaa taccgtgctc tcttctttgt 1800caatctctgg ttttcatcat tattatatta tatatgggaa taccgtgctc tcttctttgt 1800

ccttcaccaa aactagtttc acgcctacac aacatgatta gagcctcttc atttttttaa 1860ccttcaccaa aactagtttc acgcctacac aacatgatta gagcctcttc atttttttaa 1860

tctcatcctt taagtgtatt tctatttcta ttggctattg ggacaagggg aaggtggtgc 1920tctcatcctt taagtgtatt tctatttcta ttggctattg ggacaagggg aaggtggtgc 1920

ttcttaggaa cttgagctgt tttccatctt ttgagaccca tgctttgtct ctctcatttt 1980ttcttaggaa cttgagctgt tttccatctt ttgagaccca tgctttgtct ctctcatttt 1980

taattctggg tctctttctt ctcttgtcct gattttttaa atgtgcttct tttttgcttc 2040taattctggg tctctttctt ctcttgtcct gattttttaa atgtgcttct tttttgcttc 2040

ttacaaccac cctctaaacc attcatcatg cttggtttgc ttttgcttct cctttcacag 2100ttacaaccac cctctaaacc attcatcatg cttggtttgc ttttgcttct cctttcacag 2100

gtttcaatca cgcaaaacaa tgctgcaatg atgctgtact tggagtttct tctgtgaccc 2160gtttcaatca cgcaaaacaa tgctgcaatg atgctgtact tggagtttct tctgtgaccc 2160

cttttttcct tccttcaaca atcaacccac cagagaaaag tgtctcagat tttgagacta 2220cttttttcct tccttcaaca atcaacccac cagagaaaag tgtctcagat tttgagacta 2220

ctttcaactt tcaaaacaaa gtggatggga tcttcatctt atataaccac acatcaatca 2280ctttcaactt tcaaaacaaa gtggatggga tcttcatctt atataaccac acatcaatca 2280

tttgtgctac ttctccaatt ttctttagag atgaaatgaa gagctaagca gacaagacaa 2340tttgtgctac ttctccaatt ttctttagag atgaaatgaa gagctaagca gacaagacaa 2340

gtttatttgt ttgttgctg atg ggt ctt ggg ttc aag atg cag cag age cac 2392gttatttgt ttgttgctg atg ggt ctt ggg ttc aag cg cag age cac 2392

Met Gly Leu Gly Phe Lys Met Gln Gln Ser His1 5 10Met Gly Read Gly Phe Lys Met Gln Gln Being His1 5 10

cac cct gtg gct ttg aag tta cat gag caa gct ggg age cag aga aag 2440His Pro vai Ala Leu Lys Leu His Glu Gln Ala Gly Ser Gln Arg Lys15 20 25cac cct gtg gtc ttg aag tta cat gag caa gct ggg age cag aga aag 2440His Pro will Ala Leu Lys Leu His Glu Gln Ala Gly Ser Gln Arg Lys15 20 25

ttc act ttc att cag aac ttc aga aac tgg ttt cta ccc ctt ctg ttt 2488Phe Thr Phe Ile Gln Asn Phe Arg Asn Trp Phe Leu Pro Leu Leu Phettc act ttc att cag aac ttc aga aac tgg ttt cta ccc ctt ctg ttt 2488Phe Thr Phe Ile Gln Asn Phe Arg Asn Trp Phe Leu Pro Leu Phe

30 35 4030 35 40

gta tgg ttc att gtt atg gct gea ttt ggt gcc tgc ate tac cat aaa 2536vai Trp Phe Ile vai Met Ala Ala Phe Gly Ala Cys Ile Tyr His Lys45 50 55gta tgg ttc att gtt atg gct gea ttt ggt gcc tgc until tac cat aaa 2536vai Trp Phe Ile goes Met Wing Ala Phe Gly Ala Cys Ile Tyr His Lys45 50 55

atg gat gct gaa act aaa gtc aga agg aaa gag gtg ctg ggt age ctc 2584Met Asp Ala Glu Thr Lys vai Arg Arg Lys Glu Val Leu Gly Ser Leu60 65 70 75atg gat gct gaa act aaa gtc aga agg aaa gag gtg ctg ggt age ctc 2584Met Asp Ala Glu Thr Lys goes Arg Arg Lys Glu Val Leu Gly Ser Leu60 65 70 75

tgt gat caa agg gct aga atg cta caa gac caa ttc agt gtc agt gtc 2632Cys Asp Gln Arg Ala Arg Met Leu Gln Asp Gln Phe ser Val ser Valtgt gat caa agg gct aga atg cta caa gac caa ttc agt gtc agt gtc 2632Cys Asp Gln Arg Ala Arg Met Leu Gln Asp Gln Phe Be Val Be Val

80 85 9080 85 90

aac cat gtc cat gcc ctt gcc ate ctt gtt tca acc ttc cat tac tac 2680Asn His Val His Ala Leu Ala Ile Leu vai Ser Thr Phe His Tyr Tyr95 100 105aac cat gtc cat gcc ctt gcc until ctt gtt tca acc ttc cat tac tac 2680Asn

aga aat act tca gcc att gac cag gtttgtgctt gattttcctt tccttgaagc 2734Arg Asn Thr ser Ala Ile Asp Gln110 115aga aat act tca gcc att gac cag gtttgtgctt gattttcctt tccttgaagc 2734Arg Asn Thr Be Ala Ile Asp Gln110 115

attttttagt tggaggctca atttcttttt ctgatttgat tctggcctta aaaattagaa 2794tcaattgtag aaggatttcc aaacatgccc attttggaaa ttggtgcatc tgatagtatc 2854atgtttagat cagtttcttt ttcctcagaa ttgattttgg gcttaaaatc aattgtggaa 2914ggatattcat tagtaatttg gatattgttg catcatatgg ttctatctag ttacatcatt 2974tttttccact ctgattgcat gtatctttct cctgttcttt tccctatcag gaa acc 3030attttttagt tggaggctca atttcttttt ctgatttgat tctggcctta aaaattagaa 2794tcaattgtag aaggatttcc aaacatgccc attttggaaa ttggtgcatc tgatagtatc 2854atgtttagat cagtttcttt ttcctcagaa ttgattttgg gcttaaaatc aattgtggaa 2914ggatattcat tagtaatttg gatattgttg catcatatgg ttctatctag ttacatcatt 2974tttttccact ctgattgcat gtatctttct cctgttcttt tccctatcag gaa acc 3030

Glu ThrGlu thr

ttt gea gaa tac acg gcc agg aca gea ttt gaa cgg cca tta atg agt 3078Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe Glu Arg Pro Leu Met Ser120 125 130999 9*9 9cc tat gca cag aga gtg gtt cac tca gag aga gaa aga ttt 3126Gly Val Ala Tyr Ala Gln Arg Val Val His Ser Glu Arg Glu Arg Phettt gea gaa tac acg gcc agg aca gea ttt gaa cgg cca tta atg agt 3078Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe Glu Arg Pro Leu Met Ser120 125 130999 9 * 9 9cc tat gca cag aga gtg gtt cac tca gag aga gaa aga ttt 3126Gly Val Wing Tyr Wing Gln Arg Val Val His Be Glu Arg Glu Arg Phe

135 140 145135 140 145

gag aag caa cat ggg tgg gtt ata aag aca atg gaa aga gtg cct tca 3174Glu Lys Gln His Gly Trp vai Ile Lys Thr Met Glu Arg Val Pro Ser150 155 160 165gag aag caa cat ggg tgg gtt ata aag aca atg gaa aga gtg cct tca 3174Glu Lys Gln His Gly Trp goes Ile Lys Thr Met Glu Arg Val Pro Ser150 155 160 165

ggg gtt agg gat gag tat gca gca gtg ata ttt gca cag gaa act gtc 3222Gly Val Arg Asp Glu Tyr Ala Ala vai Ile Phe Ala Gln Glu Thr Val170 175 180ggg gtt agg gat gag tat gca gca gtg ata ttt gca cag gaa act gtc 3222Gly Val Arg Asp Glu Tyr Wing Wing Go Ile Phe Wing Gln Glu Thr Val170 175 180

tct tac ctt gaa tct att gat atg atg tct ggg gag gtaaatgtca 3268tct tac ctt gaa tct att gat atg tgt ggg gag gtaaatgtca 3268

Ser Tyr Leu Glu Ser Ile Asp Met Met Ser Gly GluSer Tyr Leu Glu Ser Ile Asp Met Met Ser Gly Glu

185 190185 190

acacttgtga attaattgta aaactcagaa gctactcaga gaagctcttc cccagaattg 3328gttctgcctt tagaataaat tgtacatgga tttgaccaca ttttctcatt tgcatgatgc 3388ag gag gac cga gag aac att ttg agg gct aga gcc act ggg aaa gct 3435Glu Asp Arg Glu Asn Ile Leu Arg Ala Arg Ala Thr Gly Lys Ala195 200 205gag gac cg gg gg lg gg gg lg gg gg gg gg lg gg gg gg lg gg gg lg lg gg lg gg gg gg

gtt ctg act age cct ttc aga ctg ctg gat tct cat cat ctt ggc gtg 3483vai Leu Thr Ser Pro Phe Arg Leu Leu Asp Ser His His Leu Gly Val210 215 220gtt ctg act age cct ttc aga ctg cat ctg gat tct cat cat ctt ggc gtg 3483vai Leu Thr Be Pro Phe Arg Leu Asp Ser His His Leu Gly Val210 215 220

gtt cta aca ttt cct gtt tat aaa tct aag ctc cct cca gag cca acg 3531Val Leu Thr Phe Pro Val Tyr Lys Ser Lys Leu Pro Pro Glu Pro Thr225 230 235 240gtt cta aca ttt cct gtt tat aaa tct aag ctc cct cca gag cca acg 3531Val Read Thr Phe Pro Val Tyr Lys Ser Lys Leu Pro Pro Glu Pro Thr225 230 235 240

acg gaa gag gtc att aaa gcc ata gca gg gtatgtcctc atttcacttt 3580acg gaa gag gtc att aaa gcc ata gca gg gtatgtcctc atttcacttt 3580

Thr Glu Glu vai lie Lys Ala Ile Ala GlyThr Glu Glu will lie Lys Wing Ile Wing Gly

245245

tcttgccaaa accagacttc tatttggttg tgtttccgta ggctatgact gatatgtagt 3640ttcaactcag ttagactata atataaaccc ttcatgtggc ccttacccaa cagcttaagc 3700ttttgggata attggttgct tgacaaactc cttccgtaga aaacttggtt agctttggtt 3760ctatgtgggc tttatgtttt ccctgagctt atgtaatagc atgatgtgtt taatgtactt 3820tttaatggaa acag a tat att gga gga tcc ttt gat gtt gag tcc ctt gtg 3871Tyr lie Gly Gly Ser Phe Asp Val Glu Ser Leu Val255 260tcttgccaaa accagacttc tatttggttg tgtttccgta ggctatgact gatatgtagt 3640ttcaactcag ttagactata atataaaccc ttcatgtggc ccttacccaa cagcttaagc 3700ttttgggata attggttgct tgacaaactc cttccgtaga aaacttggtt agctttggtt 3760ctatgtgggc tttatgtttt ccctgagctt atgtaatagc atgatgtgtt taatgtactt 3820tttaatggaa acag tat att gga gga tcc ttt gat GTT gag tcc ctt gtg 3871Tyr lie Gly Gly Ser Phe Asp Val Glu Ser Leu Val255 260

gag aat tta ctt ggt caa ctt gct ggt aac caa gca att ttg gtg aag 3919Glu Asn Leu Leu Gly Gln Leu Ala Gly Asn Gln Ala Ile Leu Val Lys265 270 275gag aat tta ctt ggt caa ctt gct ggt aac caa gca att ttg gtg aag 3919Glu Asn Leu Leu Gly Gln Leu Ala Gly Asn Gln Ala Ile Leu Val Lys265 270 275

gta tat gat ata aca aac tct age gac ccc cta ate atg tat ggc age 3967Val Tyr Asp Ile Thr Asn Ser Ser Asp Pro Leu Ile Met Tyr Gly Sergta tat gat ata aca aac tct age gac ccc cta until atg tat ggc age 3967Val Tyr Asp Ile Thr Asn Ser Ser Asp Pro Leu Ile Met Tyr Gly Ser

280 285 290280 285 290

caa tat gaa gag ggt gat atg tct ctt gtc cat gaa agt aag ctt gat 4015Gln Tyr Glu Glu Gly Asp Met Ser Leu vai His Glu Ser Lys Leu Asp295 300 305 310cat gaa gag ggt gat atg tct ctt gtc cat gaa agt aag ctt gat 4015Gln Tyr Glu Glu Gly Asp Met Ser Leu Goes His Glu Ser Lys Leu Asp295 300 305 310

ttt gga gat cca tac agg aaa cat cac atg ate tgt ag gtgggtgctt 4063Phe Gly Asp Pro Tyr Arg Lys His His Met Ile Cys Arg315 320ttt gga gat cca tac agg aaa cat cac atg till tgt ag gtgggtgctt 4063Phe Gly Asp Pro Tyr Arg Lys His Met Ile Cys Arg315 320

ctagttattg ttgtagtaca ttttctgagt ttcagtggtt tatcaattat cagcagattc 4123ttatgatcaa tttttttaac ag a tat cac caa cag gca cca aca aat tgg 4173ctagttattg ttgtagtaca ttttctgagt ttcagtggtt tatcaattat cagcagattc 4123ttatgatcaa tttttttaac ag to tat ca ca ca cag gca cca ac a a tgg 4173

Tyr His Gln Gln Ala Pro Thr Asn Trp325 330Tyr His Gln Gln Pro Wing Thr Asn Trp325 330

ata gca tat acc acg gca ttc cta ttc ttt gtg att ctt tgt tta gtg 4221Ile Ala Tyr Thr Thr Ala Phe Leu Phe Phe Val Ile Leu Cys Leu vai335 340 345gta tat acc acc acg gca ttc cta ttc ttt gtg att ctt tgt tta gtg 4221Ile Wing Tyr Thr Thr Wing Phe Leu Phe Val Ile Leu Cys Leu vai335 340 345

ggt tac att tta tat gct gct gga act cac att gtc aag gta gaa gat 4269Gly Tyr Ile Leu Tyr Ala Ala Gly Thr His Ile Val Lys Val Glu Aspggt tac att tta tat gct gct gga act cac att gtc aag gta gaa gat 4269Gly Tyr Ile Leu Tyr Wing Ally Gly Thr His Ile Val Lys Val Glu Asp

350 355 360350 355 360

gat tac aat gca atg cag gat tta aaa gtc aaa gca gaa gca gct gat 4317Asp Tyr Asn Ala Met Gln Asp Leu Lys Val Lys Ala Glu Ala Ala Asp365 370 375 380gat tac aat gca atg cag gat tta aaa gtc aaa gca gaa gca gct gat 4317Asp Tyr Asn Ala Met Gln Asp Leu Lys Val Lys Ala Glu Ala Asp365 370 375 380

att gcc aag tca cag gtacttttca tgacatgtta gcactgttcg ttatttcctt 4372Ile Ala Lys Ser Gln385att gcc aag tca cag gtacttttca tgacatgtta gcactgttcg ttatttcctt 4372Ile Ala Lys Ser Gln385

gaattgcata ctgatcacta gaaactgaaa atttgttatt aatgtcag ttt cta gct 4429Phe Leu Alaacc gtc tct cat gaa att aga act ccc atg aat gga att tta g 4472gaattgcata ctgatcacta gaaactgaaa atttgttatt aatgtcag ttt cta gct 4429Phe Leu Alaacc gtc tct cat gaa att aga act ccc atg aat gga att tta g 4472

Thr Val Ser His Glu Ile Arg Thr Pro Met Asn Gly Ile LeuThr Val Be His Glu Ile Arg Thr Pro Met Asn Gly Ile Leu

390 395 400390 395 400

gtaactttaa gattctctct cgttctcttt ccactgaaaa gcaacacatg ctttcattcc 4532atacctgata ctttcccatt agtgatgcta tcgttaaact ccttgtcact gtag ga 4588Glygtaactttaa gattctctct cgttctcttt ccactgaaaa gcaacacatg ctttcattcc 4532atacctgata ctttcccatt agtgatgcta tcgttaaact ccttgtcact gtag ga 4588Gly

atg ctt ggt ctg ctt tta cgc aca gaa ttg agt tca aca caa aga gac 4636Met Leu Gly Leu Leu Leu Arg Thr Glu Leu Ser Ser Thr Gln Arg Asp 405 410 415atg ctt ctt tta ggt ctg cgc gaa ttg aca aca agt tca caa aga GAC 4636Met Leu Gly Arg Leu Leu Leu Glu Thr Leu Gln Arg Ser Ser Thr Asp 405 410 415

tat gct cag act gct caa gca tgt ggg aag gca cta ata gca tta ata 4684Tyr Ala Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu Ile Ala Leu lie420 425 430 435tat gct cag act gct caa gca tgt ggg aag gca cta ata gca tta ata 4684Tyr Ala Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu Ile Ala Leu lie420 425 430 435

aat gag gtg ctt gac cga gct aaa att gaa gca ggc aaa tta gag cta 4732Asn Glu Val Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys Leu Glu Leuaat gag gtg ctt gac cga gct aaa att gaa gca ggc aaa tta gag cta 4732Asn Glu Val Leu Asp Arg Wing Lys Ile Glu Wing Gly Lys Leu Glu Leu

440 445 450440 445 450

gaa gca gtt cca ttt gac ctt cgt tcc ata ctt gac gat gtc ctt tct 4780Glu Ala Val Pro Phe Asp Leu Arg Ser Ile Leu Asp Asp vai Leu Ser455 460 465gaa gca gtt cca ttt gac ctt cgt tcc ata ctt gac gat gtc ctt tct 4780Glu Val Pro Phe Wing Asp Leu Arg Ser Ile Leu Asp Asp Go Leu Ser455 460 465

ctt ttt tct gag aag tca aga cac aaa ggc tta gag gtacgtttag 4826ctt ttt tct gag aag tca aga cac aaa ggc tta gag gtacgtttag 4826

Leu Phe Ser Glu Lys Ser Arg His Lys Gly Leu GluLeu Phe Be Glu Lys Be Arg His Lys Gly Leu Glu

470 475470 475

tcattgctaa atctgttgtg aagtcctgta caagtggcgt aatttcatag tcctatttcc 4886ttttcttcaa ttgatgatct tatttcatat tcctctcgtg tttctctctt tctatgttgc 4946catgtcgttg ggcggtgatg ttcctactct gatcccaaat tcctgatgtg caattattcc 5006gacttggact tcaatggttt gggaagtata g ctg gca gtg ttt gtt tct gac 5058tcattgctaa atctgttgtg aagtcctgta caagtggcgt aatttcatag tcctatttcc 4886ttttcttcaa ttgatgatct tatttcatat tcctctcgtg tttctctctt tctatgttgc 4946catgtcgttg ggcggtgatg ttcctactct gatcccaaat tcctgatgtg caattattcc 5006gacttggact tcaatggttt gggaagtata g ctg gca gtg gtt tct gac ttt 5058

Leu Ala vai Phe Val Ser Asp480 485Leu Wing goes Phe Val Ser Asp480 485

aaa gtt ccg gat ata gtt atg ggc gat cct ggg cga ttc aga caa ata 5106Lys Val Pro Asp Ile vai Met Gly Asp Pro Gly Arg Phe Arg Gln lie490 495 500aaa gtt ccg gat ata gtt atg ggc gat cct ggg cga ttc aga caa ata 5106Lys Val Pro Asp Ile Goes Met Gly Asp Pro Gly Arg Phe Arg Gln lie490 495 500

gtg aca aat ctt gtt gga aac tct gtt aag gttagtggaa ttttcaaact 5156Val Thr Asn Leu Val Gly Asn Ser Val Lysgtg aca aat ctt gtt gga aac tct gtt aag gttagtggaa ttttcaaact 5156Val Thr Asn Leu Val Gly Asn Ser Val Lys

505 510505 510

ttatttgcct aatgttgtgt gcaagttgtg tgttggaaat gcgtcctttt aacgttataa 5216aatcgtacaa gttcgtattc tccattgtat acaataactt attagcaaag tacttgttga 5276tatcattact gattaacttt aatatcttgc ag ttc act gag cga ggt cat ata 5329ttatttgcct aatgttgtgt gcaagttgtg tgttggaaat gcgtcctttt aacgttataa 5216aatcgtacaa gttcgtattc tccattgtat acaataactt attagcaaag tacttgttga 5276tatcattact gattagg cggtgatta

Phe Thr Glu Arg Gly His Ile515Phe Thr Glu Arg Gly His Ile515

ttt gtt aaa gtc cat tta gct gaa aaa aga cag tgc aca atg aat gga 5377Phe Val Lys Val His Leu Ala Glu Lys Arg Gln Cys Thr Met Asn Gly520 525 530 535ttt gtt aaa gtc cat tta gct gaa aaa aga cag tgc aca atg aat gga 5377Phe Val Lys Val His Leu Gla Lys Arg Gln Cys Thr Met Asn Gly520 525 530 535

aaa tgt gag act ttt cta aat gga ggc tgt gat gat gtt ttg cat gta 5425Lys Cys Glu Thr Phe Leu Asn Gly Gly Cys Asp Asp vai Leu His vaiaaa tgt gag act ttt cta aat gga ggc tgt gat gat gtt ttg cat gta 5425Lys Cys Glu Thr Phe Leu Asn Gly Gly Cys Asp Asp will Leu His will

540 545 550540 545 550

tct ggc agt tat aat ttg aaa acc ctt agt gga tat gaa gcc gct gat 5473Ser Gly Ser Tyr Asn Leu Lys Thr Leu Ser Gly Tyr Glu Ala Ala Asp555 560 565tct ggc agt tat aat ttg aaa acc ctt agt gga tat gaa gcc gct gat 5473Ser Gly Ser Tyr Asn Leu Lys Thr Leu Ser Gly Tyr Glu Wing Asp555 560 565

gaa cgg aac age tgg gat aat ttt aag cat cat att gct gac gaa gaa 5521Glu Arg Asn Ser Trp Asp Asn Phe Lys His His Ile Ala Asp Glu Glu570 575 580cat cgg aac age tgg gat aat ttt aag cat cat att gct gac gaa gaa 5521Glu Arg Asn Ser Trp Asp Asn Phe Lys His Ile Ala Asp Glu Glu570 575 580

ttt ttc ttt gat gct tcg gtt aaa aag ttg gcc tct agt gaa tct tat 5569Phe Phe Phe Asp Ala Ser Val Lys Lys Leu Ala Ser Ser Glu Ser Tyrttt ttc ttt gat gct tcg gtt aaa aag ttg gcc tct agt gaa tct tat 5569Phe Phe Phe Asp Ala Ser Val Lys Lys Leu Ala Ser Ser Glu Ser Tyr

585 590 595585 590 595

gag caa gtc acc ttg atg gtc age gtg gag gac act gga att ggg att 5617Glu Gln vai Thr Leu Met Val Ser Val Glu Asp Thr Gly lie Gly Ile600 605 610 615gag caa gtc acc ttg atg gtc age gtg gag gac act gga att ggg att 5617Glu Gln will Thr Leu Met Val Ser Val Glu Asp Thr Gly lie Gly Ile600 605 610 615

tct ttc tct gcc caa gat agt att ttc atg cct ttt gtg cag gct gac 5665Ser Phe Ser Ala Gln Asp Ser Ile Phe Met Pro Phe Val Gln Ala Asptct ttc tct gcc caa gat agt att ttc atg cct ttt gtg cag gct gac 5665Ser Phe Ser Ala Gln Asp Ser Ile Phe Met Pro Phe Val Gln Ala Asp

620 625 630620 625 630

age tca acc tct cga aac tat ggg ggt acc ggg ate ggc ttg agt ate 5713ser ser Thr ser Arg Asn Tyr Gly Gly Thr Gly Ile Gly Leu ser lie635 640 645age tca acc tct cga aac tat ggg ggt acc ggg until ggc ttg agt until 5713ser be Thr be Arg Asn Tyr Gly Gly Thr Gly Ile Gly Read ser lie635 640 645

agt aag tgc ttg gtt gaa ctg atg ggc ggt cag ata aac ttc ata age 5761Ser Lys Cys Leu vai Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Seragt aag tgc ttg gtt gaa ctg atg ggc ggt cag ata aac ttc ata age 5761Ser Lys Cys Leu goes Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Ser

650 655 660650 655 660

cga ccc cag gtc ggg age acg ttt tca ttc act gea gat ttc gga aca 5809Arg Pro Gln Val Gly Ser Thr Phe ser Phe Thr Ala Asp Phe Gly Thr 665 670 675cga ccc cag gtg ggg age acg ttt tca ttc act gea gat ttc gga aca 5809Arg Pro Gln Val Gly Be Thr Phe Be Phe Thr Wing Asp Phe Gly Thr 665 670 675

ttt aag aaa aac tca aca act gac atg aag aaa ctt aac ttt gaa gat 5857Phe Lys Lys Asn Ser Thr Thr Asp Met Lys Lys Leu Asn Phe Glu Asp680 685 690 695att aaa aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 5857Phe

cta cct tet agt ttt aga ggt ctt aaa gcc att gtg gtt gat gga aaa 5905Leu Pro Ser Ser Phe Arg Gly Leu Lys Ala Ile Val Val Asp Gly Lyscta cct tet agt ttt aga ggt ctt aaa gcc att gtg gtt gat gga aaa 5905Leu Pro Be Ser Phe Arg Gly Leu Lys Ala Ile Val Val Asp Gly Lys

700 705 710700 705 710

cct gtt aga gct gea gtg act aga tac cat ttg aag aga cta ggg ata 5953Pro Val Arg Ala Ala vai Thr Arg Tyr His Leu Lys Arg Leu Gly Ile715 720 725cct gtt aga gct gea gtg act aga tac cat ttg aag aga cta ggg ata 5953Pro Val Arg Wing Ward Go Thr Arg Tyr His Leu Lys Arg Leu Gly Ile715 720 725

caa gct aaa gtt gea att age ate aat aag gct gtt tet tta tgt ggg 6001Gln Ala Lys vai Ala Ile Ser Ile Asn Lys Ala Val Ser Leu Cys Glycaa gct aaa gtt gea att age till aat aag gct gtt tet tta tgt ggg 6001Gln Ala Lys goes Ala Ile Ser Ile Asn Lys Ala Val Ser Leu Cys Gly

730 735 740730 735 740

aaa aat ggt tet ttg acc tcg gc gtaagtcttt aattaacctt tttggtttca 6054Lys Asn Gly Ser Leu Thr Ser Ala20 745 750aaa aat ggt tet ttg acc tcg gc gtaagtcttt aattaacctt tttggtttca 6054Lys Asn Gly Ser Leu Thr Ser Ala20 745 750

attatgtaga aatgtattga atgttatgat aatcagtagc attatcaact tttagtaatt 6114gttcttaaca tatgctaata gtcatatctt tctataatac tacaatactg tagccatata 6174atatctttcc tgtattggag tgagttttca aatgtttttc tgtgatattt tggaagttat 6234cttcagtttg agaactcatt tgtcattttt gcattttgtt attggatatt tggatggatc 6294tttacaaagg atgtgtggat tttgacttgt tatacacatt tcttctccat tttatattgt 6354ttgtgttatt ctttttactc ataaagaaat ttagaaactg cattgactgg ttctttttaa 6414ttacttacag atattgacat tgatattttt tgtaaatgct gtcttgacat ggtttaatta 6474cttacagact aggtttttct ttccttttct aacatgcata tccatttact tttttgacca 6534accaacatcc tcatgagtca tgacatgttg atgatttata tggttgactt gagactattt 6594agacattaaa taaccgcaaa ttccatgttg tttgtgtgtt tggttccctg ttgggtaatc 6654tcagaatcaa ttatgataga gtaaaatcaa ttttggatga gatgtgtggg tgtcattttg 6714taaacctaaa cccaaaatca attctgctat aagetagaga gagtagttga acataatcaa 6774ttgtgagatt ttgcaagtgg attgcacaac attgccctat gaaaatcact ttttgttcac 6834aaaatttatc taaacataca taacttcatt ttcaaccttt actataatca attttacaat 6894aattaatttt acccaaaatc aattgtgaca atgagtttcc aaacacacac ttaaagacta 6954ccatttgcag aaaatatgtg atagaagact tatgtttatg tagtgtgttt cagttcattc 7014actgatttaa actactcgga ttttgcag a tta ttt cag cct gat att att ttt 7067attatgtaga aatgtattga atgttatgat aatcagtagc attatcaact tttagtaatt 6114gttcttaaca tatgctaata gtcatatctt tctataatac tacaatactg tagccatata 6174atatctttcc tgtattggag tgagttttca aatgtttttc tgtgatattt tggaagttat 6234cttcagtttg agaactcatt tgtcattttt gcattttgtt attggatatt tggatggatc 6294tttacaaagg atgtgtggat tttgacttgt tatacacatt tcttctccat tttatattgt 6354ttgtgttatt ctttttactc ataaagaaat ttagaaactg cattgactgg ttctttttaa 6414ttacttacag atattgacat tgatattttt tgtaaatgct gtcttgacat ggtttaatta 6474cttacagact aggtttttct ttccttttct aacatgcata tccatttact tttttgacca 6534accaacatcc tcatgagtca tgacatgttg atgatttata tggttgactt gagactattt 6594agacattaaa taaccgcaaa ttccatgttg tttgtgtgtt tggttccctg ttgggtaatc 6654tcagaatcaa ttatgataga gtaaaatcaa ttttggatga gatgtgtggg tgtcattttg 6714taaacctaaa cccaaaatca attctgctat aagetagaga gagtagttga acataatcaa 6774ttgtgagatt ttgcaagtgg attgcacaac attgccctat gaaaatcact ttttgttcac 6834aaaatttatc taaacataca taacttcatt ttcaaccttt actataatca attttacaat 6894aattaatttt acccaaaat c aattgtgaca atgagtttcc aaacacacac ttaaagacta 6954ccatttgcag aaaatatgtg atagaagact tatgtttatg tagtgtgttt cagttcattc 7014actgatttaa actactcgga ttttgcag a tta ttt cag cct gat att ttt 7067

Leu Phe Gln Pro Asp Ile Ile Phe755Read Phe Gln Pro Asp Ile Ile Phe755

gtt gag aag gac tct tgg gtt tct gga gag gat ggt ggt ate ttc aat 7115vai Glu Lys Asp Ser Trp vai Ser Gly Glu Asp Gly Gly Ile Phe Asn 760 765 770 775gt gag aag gac tct tgg gtt tct gga gag gat ggt ggt till ttc aat 7115vai Glu Lys Asp Be Trp Will Be Gly Glu Asp Gly Gly Ile Phe Asn 760 765 770 775

gcg ttt aag atg cct caa atg ate ctt ctt gea acc aat ate tgt aac 7163Ala Phe Lys Met Pro Gln Met Ile Leu Leu Ala Thr Asn Ile Cys Asngcg ttt aag atg cct hunt atg until ctt ctt gea acc aat until tgt aac 7163Ala Phe Lys Met Pro Gln Met Ile Leu Leu Thr Asn Ile Cys Asn

780 785 790780 785 790

gct gaa ttt gat aaa gcc aaa gct gea ggt ttc agt gat aca gtg ate 721115 Ala Glu Phe Asp Lys Ala Lys Ala Ala Gly Phe Ser Asp Thr vai Ile795 800 805gt gaa ttt gat aaa gcc aaa gct gea ggt ttc agt gat aca gtg till 721115 Ala Glu Phe Asp Lys Ala Lys Ala Gly Phe Ser Asp Thr Go Ile795 800 805

atg aag cca ctg aga gct agt atg ctg gct gct tgt ctt cag caa gtt 7259Met Lys Pro Leu Arg Ala Ser Met Leu Ala Ala Cys Leu Gln Gln vai810 815 820atg aag cca ctg aga gct agt atg ctg gct gct tgt ctt cag caa gtt 7259Met Lys Pro Leu Arg Ala Ser Met Leu Ala Cys Leu Gln Gln vai810 815 820

ttc ggg act ggc aag acg agg cag ttt ggg aaa gac atg tcg aat ggt 7307Phe Gly Thr Gly Lys Thr Arg Gln Phe Gly Lys Asp Met Ser Asn Glyttc ggg act ggc aag acg agg cag ttt ggg aaa gac atg tcg aat ggt 7307Phe Gly Thr Gly Lys Thr Arg Gln Phe Gly Lys Asp Met Ser Asn Gly

825 830 835825 830 835

tct tca gta cga age ctt ctt tgc gga aag aaa ate tta gtg gtt gat 7355Ser Ser Val Arg Ser Leu Leu Cys Gly Lys Lys Ile Leu Val Val Asp 840 845 850 855tct tca gta cga age ctt ctt tgc gga aag aaa till tta gtg gtt gat 7355Ser Ser Val Arg Ser Leu Cys Gly Lys Lys Ile Le Val Val Asp 840 845 850 855

gat aat ttg gtg aac cga agg gtc gcc gcc ggc gcg ttg aaa aac ttt 7403Asp Asn Leu Val Asn Arg Arg vai Ala Ala Gly Ala Leu Lys Asn Phegat aat ttg gtg aac cga agg gtc gcc gcc gcc gcg ttg aaa aac ttt 7403Asp Asn Leu Val Asn Arg Arg goes Ala Ala Gly Ala Leu Lys Asn Phe

860 865 870860 865 870

gga gct gat gtc aaa tgt gea gea agt ggc aaa gct gct ctt gaa atg 7451 Gly Ala Asp Val Lys Cys Ala Ala Ser Gly Lys Ala Ala Leu Glu Met875 880 885gga gct gat gtc aaa tgt gea gea agt ggc aaa gct gct ctt gaa atg 7451 Gly Ala Asp Val Lys Cys Ala Ser Gly Lys Ala Leu Glu Met875 880 885

ctt caa tat cct cac gat ttc gat gct tgc ttc atg gat att caa atg 7499Leu Gln Tyr Pro His Asp Phe Asp Ala Cys Phe Met Asp Ile Gln Metctt caa tat cct cac gat ttc gat gct tgc ttc atg gat att caa atg 7499Leu Gln Tyr Pro His Asp Phe Asp Ala Cys Phe Met Asp Ile Gln Met

890 895 900890 895 900

cca gaa atg gat gg gtatgcttac tggcactgac taatacatgt tttttgccaa 7553Pro Glu Met Asp Gly 905cca gaa atg gat gg gtatgcttac tggcactgac taatacatgt tttttgccaa 7553Pro Glu Met Asp Gly 905

cttaatatat tactctttca atattcgttg tgttattaga agatcatata gattaattta 7613taaattttct tttagcaaaa ccttatcaat taagtgtgta gaaaagtcag tctcacatta 7673tggtcaaata agtgttaggg caagcttcac ctcaaagcta gctatttggg tagatttagg 7733cctaacccga attctaagat ggtatcagag tctatcctag atctttttat tggaaaccac 7793ccgtatatga gcaactcgta gatattcatt cttgaaagtt gcacgctcca tatgtccatt 7853cctaggtgcg agagagaagt ctcactttga ctagagatat gattaaaaaa atatttataa 7913agggttgagc aatcctcacc tcagagctaa gcttttgggg taaagttagg cctaactcga 7973actctaataa agtgtttagc tggtgtgtca actgtcaata tgaaatcttt tgcaatttac 8033tatgcattca cttacctact ttattgaagc ttattgacaa tttgtgcaga agcatcatta 8093attaggaaca tgttagctat acaagttatg atgtttttgt atagcatatc atgttccaac 8153cttccaataa caaaatatgt ggttcaagtg tgagaatata taggttaaac aataaagtat 8213tgagttaaca gaaatctaaa cacacgctgt cactagctct tcatattgag acatgcatgg 8273gatttgacaa aacatctgaa taaatatttg cag g ttt gag gcc act cgt cga 8325cttaatatat tactctttca atattcgttg tgttattaga agatcatata gattaattta 7613taaattttct tttagcaaaa ccttatcaat taagtgtgta gaaaagtcag tctcacatta 7673tggtcaaata agtgttaggg caagcttcac ctcaaagcta gctatttggg tagatttagg 7733cctaacccga attctaagat ggtatcagag tctatcctag atctttttat tggaaaccac 7793ccgtatatga gcaactcgta gatattcatt cttgaaagtt gcacgctcca tatgtccatt 7853cctaggtgcg agagagaagt ctcactttga ctagagatat gattaaaaaa atatttataa 7913agggttgagc aatcctcacc tcagagctaa gcttttgggg taaagttagg cctaactcga 7973actctaataa agtgtttagc tggtgtgtca actgtcaata tgaaatcttt tgcaatttac 8033tatgcattca cttacctact ttattgaagc ttattgacaa tttgtgcaga agcatcatta 8093attaggaaca tgttagctat acaagttatg atgtttttgt atagcatatc atgttccaac 8153cttccaataa caaaatatgt ggttcaagtg tgagaatata taggttaaac aataaagtat 8213tgagttaaca gaaatctaaa cacacgctgt cactagctct tcatattgag acatgcatgg 8273gatttgacaa aacatctgaa taaatatttg g cag gag ttt gcc cgt cga act 8325

Phe Glu Ala Thr Arg Arg 910Phe Glu Wing Thr Arg Arg 910

att cgg atg atg gaa aga gag gca agt gag cag ctg aaa agt gaa tct 8373Ile Arg Met Met Glu Arg Glu Ala Ser Glu Gln Leu Lys ser Glu Ser915 920 925 930att cgg atg atg gaa aga gag gca agt gag cag ctg aaa agt gaa tct 8373Ile Arg Met Met Glu Arg Glu Ala Ser Glu Gln Leu Lys Be Glu Ser915 920 925 930

ggt gaa gaa aat ggt aag aaa agt gag ttc cac atg cct ata ttg gcc 8421Gly Glu Glu Asn Gly Lys Lys Ser Glu Phe His Met Pro Ile Leu Alaggt gaa gaa aat ggt aag aaa agt gag ttc cac atg cct ata ttg gcc 8421Gly Glu Glu Asn Gly Lys Lys Ser Glu Phe His Met Pro Ile Leu Ala

935 940 945935 940 945

atg aca gct gat gta ate cat gct aca tat gat aag tgc tta aat tgt 8469Met Thr Ala Asp vai Ile His Ala Thr Tyr Asp Lys Cys Leu Asn Cys950 955 960atg aca gct gat gta until cat gct aca tat gat aag tgc tta aat tgt 8469Met Thr Ala Asp Goes Ile His Ala Thr Tyr Asp Lys Cys Leu Asn Cys950 955 960

ggg atg gat gga tac gtc tca aag cct ttt gaa gaa gag aat ctc tat 8517Gly Met Asp Gly Tyr vai Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr965 970 975caa gca gtt gca aag ttc ttc aag tcc aag cct gct tca gac tca tg 8564Gln Ala vai Ala Lys Phe Phe Lys Ser Lys Pro Ala ser Asp serggg atg gat gga tac gtc tca aag cct ttt gaa gaa gag aat ctc tat 8517Gly Met Asp Gly Tyr Will Be Lys Pro Phe Glu Glu Asn Leu Tyr965 970 975caa gca gtt gca aag tcc aag tcc cc gt tca 85 Wing Go Wing Lys Phe Phe Lys Be Lys Pro Wing Be Asp Be

980 985 990 acactgctta ttctgcagaa caggtcaacc aacttttgat tgagaaacat ttagtgttag 8624 catgtttgga tcaacttctc ccagcatcaa ttctgaaact cagaagctac tcatagaagc 8684 ttctagccag aattgatttt ggcttcagaa gctactaatg gttttatgta gagagcaaaa 8744 gttggtttcc aagatatgcg aggatccatg atgatacaca cgtctgaagt gatcatttct 8804 aaccagttga agtttcactc gacgtgattt gaatccaagt aaatgcatac cacataatta 8864 tccatcccca tcttgtgtac agattctccc aagggataag aaaatttatg taaattcaat 8924 ttttttcttt tgcatctcaa tacttccctg ttagaacttt ttccctatga ttatttccac 8984 ttttcatttt caattatatt ttttgtaaaa ttggtctcca tcctataagg tttgcttagt 9044 tttttttaat tcagatgaga aggttggtgc ttataatgtg tatacctttt tagcagtact 9104 tgtgatataa atatttgtct tatttttaga acttaggaga taataagatc gtaggagtaa 9164 atagcatttt tggagaatga tatcataatt ataatcatct acattctgca gttataaaaa 9224 aaatcaatat taatgcatag caagtagctt taagaagtat gatctattta agtattgaat 9284 tttttttcta aactaattac aatgtattat tgctgggata tgcgcttttt actcgataga 9344 tatcatgttt ggattagttt ttcttagaat ccattttgga accgaaaaaa ttatcacata 9404 atttattctt caaagttgat tttggcaatc aaggtaaaca ctcataaaag ctaaagggag 9464 ttgtttctta ttattgtgat tttagctcat tgtggttttt caccgtgtat ataaacatgc 9524 atttgatgtg acgttgactt gtgaagcacg tggctatcca ccagcaatct caagtaacac 9584 ttgtcagcac tatgttgcta ttataagtga caaggcagtg accacacccc tctatatatg 9644 cataaacttt tttattaaaa gtaacagaat caaaagggaa gttttctaca aattaattaa 9704 tggaaaatta tcagaaccag tatggagctg ttcctgctag cagggctccg gttcagagcc 9764 agagtgaaat attggaaacc accaatgttg gcggtgctac acaagtcata agaaatgatt 9824 atgatggaag aaccactgct gcagatgcta ctgttgttgg tgagagcgat ctgcagcatc 9884 accacaagaa aggaatatta gagaagatca aggagaagct tcctggaaca caccaccatc 9944 aagatcacaa atagctagct agctttgttt agtttatttt gcttctatca ttataaatgt 10004 aataataata gtatgtgtgt tgctacatgc atgtgtatgt atcgtttagt cagctagttt 10064 gttaataact tgctttcaat tttgcttcct tacaagccta tttatgcaat gtatttggtc 10124 atttcactat tggtttctaa tcaatctaaa ttagtacatg gtcctcttcc ttttagtgcc 10184 ttttaattaa agagtaaatg ttgaatattg tttttagtca ttctatttac atttttattt 10244 tttttgcttc atcggaaaat tatttacatt tttattaagt gtgtgtttag attttcgttt 10304ttcgttaaaa atttcataat taatttttta taacagtgat gtatgcagct aaggcatcct 10364 aacagaccta tctttgtatg tatggcattc tgggcgggtc ggatttattt ttagccccat 10424 aattccccac tactatacat aaaatgatac ctgctagtaa aataatttaa agtgtttcga 10484 agtcatattg gtaaattgat ttcagttaaa aatcgaagtt tgttagctac taaagacggt 10544 agcttctatg tcaaaatagt taattcctat attttataat tggatcattt tttctaaaat 10604 ttattcaaac atctcaactc actttgacta taatcacttt ataattaatt ttgttgaatt 10664 aaataaatta tattcaaatt aaatgtgata atacccactg aatgtgattg gaatgccatc 10724 agggaaatat ggaacaaaga gagctgcatg tggaagggct tgttatgtct gcaaaacaga 10784 caaagaagtt gtaatcacaa ttaggacaat tgccaaagat gtggggtcac attgacttgt 10844 cagggaaagt ctatagatcc actgtgctct taaagctaga acaattaaca taaaatttct 10904 gccaattaag gctttcactt cctatcccta tctgctgtat attttaagtt tgagtaaagg 10964 gacagttgtg gtgatgttat aatcaactgc aggctgaggg aaaagttggg ggtcaagtgg 11024 tgtctatata aacttttttg tcattattaa tctttaatct ttttggtttt gtaaaatgac 11084 agcttttgtc ttgtgctgaa agttgactgt gctagctagt aaagtcatgc tggtttctta 11144 attgattgtg gtgtttttat gcgacaactg tagacattgc aatgcaggca cacaagtcaa 11204 gaggcaatta caaacaaaat acagctatta ataattgacc tactaactac taaggaggga 11264 atttaaatgt gattttgaaa tcttatattg gatgtaaaga ggcaatactc atcagactgt 11324 aaagtgacag aacaagtaga tggtaaacaa gtagaagtag aagcagcaac aaattgagta 11384 aagaccatca atgtcaacac ttccttggag aagaatttca taagtgtcat gtgaaaataa 11444 ctagacaaaa tgtagccaaa atgaagtatg aaactcagtt attgaaacga gtgacactga 11504 ctagattcca agttattgaa ggaaccagca aaaggttaag tgacagtgaa gtgtgtgggt 11564 tgaaagagga ttgaataaaa gctgaaccaa tggactgaat aggcaaactt gacaatttcc 11624 cattagaatc tgttttaact tccttgtaca aaagtttctt cggtgaaatt tccagcattg 11684 ttggtaacat gatgagtagc tcatgaatct ggaaaccaaa cagttttgtt tgttcaagct 11744 gtagagaaga atgagaacta catgtgagca ttgttatgta ccaaaaaaac ctgtgagcat 11804 tgttgctgca atcctggatc agtacgcata gcctgagcac aaagatgttg aattgaagta 11864 gttgctctaa tatgttatga atattcaaga ttaaaacatt tttaagttat catgtgacaa 11924 tcacataact attaaattat tttaatctta actattaatc attagatcta acgatcatat 11984 tctcatataa aataattatt ctgatacaat aatatatata tatatatata tgtatgtagg 12044 ggatctgagg atgtatcata tgagaaatgt gatattatgt gagaacatga gaataaatca 12104 caaccgttag atttaatcaa cggaccagat taaaacaatt taatgctcac atgaccactt 12164 aaaaatgtca tgtgactttc ttaaaaagtc aaatgacttc atgttttaat tttgaccgtt 12224catgattaga tttaacggtc atgatttatt ctcatgttct cacataaaaa catttttcat 12284ttttctgata tgtatatttg taacataaaa cacaacataa aatactatgt aaatcagcca 12344atttcaccaa atacaatatg atgtgaaaaa tataatatta ctattgatat tcactcaaat 12404gcacaatcgg ccctaaaaaa tggtcaattt ttctggttaa tatatttctt ttttaaaaca 12464tgagtattca tgcaatatat gatttcaaaa tatataattt ctttcaacat ttaaccaatt 12524tgcaccatgt gtaatatgta agagatcaaa aattatattg agttttattt atcttttaac 12584tgatatggga aacattttta tagataattt ttattcaagt tctaagttac ttcattttta 12644tttcggcgaa gttggtgctt ttttttcttt tcaaaccgtt tgaagatggt ttcctcatgg 12704attggagttt aatccatttt tctatcatga gctc 12738<210> 2 <211> 3568 <212> DNA <213> Lotus japonicus <220> <221> 5'UTR <222> Cl). .(137) <220> <221> CDS <222> (138)..(3119) <220> <221> 3'UTR <222> (3120)..(3568) <400> 2980 985 990 acactgctta ttctgcagaa caggtcaacc aacttttgat tgagaaacat ttagtgttag 8624 catgtttgga tcaacttctc ccagcatcaa ttctgaaact cagaagctac tcatagaagc 8684 ttctagccag aattgatttt ggcttcagaa gctactaatg gttttatgta gagagcaaaa 8744 gttggtttcc aagatatgcg aggatccatg atgatacaca cgtctgaagt gatcatttct 8804 aaccagttga agtttcactc gacgtgattt gaatccaagt aaatgcatac cacataatta 8864 tccatcccca tcttgtgtac agattctccc aagggataag aaaatttatg taaattcaat 8924 ttttttcttt tgcatctcaa tacttccctg ttagaacttt ttccctatga ttatttccac 8984 ttttcatttt caattatatt ttttgtaaaa ttggtctcca tcctataagg tttgcttagt 9044 tttttttaat tcagatgaga aggttggtgc ttataatgtg tatacctttt tagcagtact 9104 tgtgatataa atatttgtct tatttttaga acttaggaga taataagatc gtaggagtaa 9164 atagcatttt tggagaatga tatcataatt ataatcatct acattctgca gttataaaaa 9224 aaatcaatat taatgcatag caagtagctt taagaagtat gatctattta agtattgaat 9284 tttttttcta aactaattac aatgtattat tgctgggata tgcgcttttt actcgataga 9344 tatcatgttt ggattagttt ttcttagaat ccattttgga accgaaaaaa ttatca vane 9404 atttattctt caaagttgat tttggcaatc aaggtaaaca ctcataaaag ctaaagggag 9464 ttgtttctta ttattgtgat tttagctcat tgtggttttt caccgtgtat ataaacatgc 9524 atttgatgtg acgttgactt gtgaagcacg tggctatcca ccagcaatct caagtaacac 9584 ttgtcagcac tatgttgcta ttataagtga caaggcagtg accacacccc tctatatatg 9644 cataaacttt tttattaaaa gtaacagaat caaaagggaa gttttctaca aattaattaa 9704 tggaaaatta tcagaaccag tatggagctg ttcctgctag cagggctccg gttcagagcc 9764 agagtgaaat attggaaacc accaatgttg gcggtgctac acaagtcata agaaatgatt 9824 atgatggaag aaccactgct gcagatgcta ctgttgttgg tgagagcgat ctgcagcatc 9884 accacaagaa aggaatatta gagaagatca aggagaagct tcctggaaca caccaccatc 9944 aagatcacaa atagctagct agctttgttt agtttatttt gcttctatca ttataaatgt 10004 aataataata gtatgtgtgt tgctacatgc atgtgtatgt atcgtttagt cagctagttt 10064 gttaataact tgctttcaat tttgcttcct tacaagccta tttatgcaat gtatttggtc 10124 atttcactat tggtttctaa tcaatctaaa ttagtacatg gtcctcttcc ttttagtgcc 10184 ttttaattaa agagtaaatg ttgaatattg tttttagtca ttctatttac atttttat tt 10244 tttttgcttc atcggaaaat tatttacatt tttattaagt gtgtgtttag attttcgttt 10304ttcgttaaaa atttcataat taatttttta taacagtgat gtatgcagct aaggcatcct 10364 aacagaccta tctttgtatg tatggcattc tgggcgggtc ggatttattt ttagccccat 10424 aattccccac tactatacat aaaatgatac ctgctagtaa aataatttaa agtgtttcga 10484 agtcatattg gtaaattgat ttcagttaaa aatcgaagtt tgttagctac taaagacggt 10544 agcttctatg tcaaaatagt taattcctat attttataat tggatcattt tttctaaaat 10604 ttattcaaac atctcaactc actttgacta taatcacttt ataattaatt ttgttgaatt 10664 aaataaatta tattcaaatt aaatgtgata atacccactg aatgtgattg gaatgccatc 10724 agggaaatat ggaacaaaga gagctgcatg tggaagggct tgttatgtct gcaaaacaga 10784 caaagaagtt gtaatcacaa ttaggacaat tgccaaagat gtggggtcac attgacttgt 10844 cagggaaagt ctatagatcc actgtgctct taaagctaga acaattaaca taaaatttct 10904 gccaattaag gctttcactt cctatcccta tctgctgtat attttaagtt tgagtaaagg 10964 gacagttgtg gtgatgttat aatcaactgc aggctgaggg aaaagttggg ggtcaagtgg 11024 tgtctatata aacttttttg tcattattaa tctttaatct g ttttggtttt taaaatgac 11084 agcttttgtc ttgtgctgaa agttgactgt gctagctagt aaagtcatgc tggtttctta 11144 attgattgtg gtgtttttat gcgacaactg tagacattgc aatgcaggca cacaagtcaa 11204 gaggcaatta caaacaaaat acagctatta ataattgacc tactaactac taaggaggga 11264 atttaaatgt gattttgaaa tcttatattg gatgtaaaga ggcaatactc atcagactgt 11324 aaagtgacag aacaagtaga tggtaaacaa gtagaagtag aagcagcaac aaattgagta 11384 aagaccatca atgtcaacac ttccttggag aagaatttca taagtgtcat gtgaaaataa 11444 ctagacaaaa tgtagccaaa atgaagtatg aaactcagtt attgaaacga gtgacactga 11504 ctagattcca agttattgaa ggaaccagca aaaggttaag tgacagtgaa gtgtgtgggt 11564 tgaaagagga ttgaataaaa gctgaaccaa tggactgaat aggcaaactt gacaatttcc 11624 cattagaatc tgttttaact tccttgtaca aaagtttctt cggtgaaatt tccagcattg 11684 ttggtaacat gatgagtagc tcatgaatct ggaaaccaaa cagttttgtt tgttcaagct 11744 gtagagaaga atgagaacta catgtgagca ttgttatgta ccaaaaaaac ctgtgagcat 11804 tgttgctgca atcctggatc agtacgcata gcctgagcac aaagatgttg aattgaagta 11864 gttgctctaa tatgttatga atattcaaga ttaaaacatt TTTA agttat catgtgacaa 11924 tcacataact attaaattat tttaatctta actattaatc attagatcta acgatcatat 11984 tctcatataa aataattatt ctgatacaat aatatatata tatatatata tgtatgtagg 12044 ggatctgagg atgtatcata tgagaaatgt gatattatgt gagaacatga gaataaatca 12104 caaccgttag atttaatcaa cggaccagat taaaacaatt taatgctcac atgaccactt 12164 aaaaatgtca tgtgactttc ttaaaaagtc aaatgacttc atgttttaat tttgaccgtt 12224catgattaga tttaacggtc atgatttatt ctcatgttct cacataaaaa catttttcat 12284ttttctgata tgtatatttg taacataaaa cacaacataa aatactatgt aaatcagcca 12344atttcaccaa atacaatatg atgtgaaaaa tataatatta ctattgatat tcactcaaat 12404gcacaatcgg ccctaaaaaa tggtcaattt ttctggttaa tatatttctt ttttaaaaca 12464tgagtattca tgcaatatat gatttcaaaa tatataattt ctttcaacat ttaaccaatt 12524tgcaccatgt gtaatatgta agagatcaaa aattatattg agttttattt atcttttaac 12584tgatatggga aacattttta tagataattt ttattcaagt tctaagttac ttcattttta 12644tttcggcgaa gttggtgctt ttttttcttt tcaaaccgtt tgaagatggt ttcctcatgg 12704attggagttt aatccatttt tctatcatga GCTC 12738 <210> 2 <211> 3568 <212> DNA <213> Lotus japonicus <220> <221> 5'UTR <222> Cl). . (137) <220> <221> CDS <222> (138) .. (3119) <220> <221> 3'UTR <222> (3120) .. (3568) <400> 2

ttcaactttc aaaacaaagt ggatgggatc ttcatcttat ataaccacac atcaatcatt 60tgtgctactt ctccaatttt ctttagagat gaaatgaaga gctaagcaga caagacaagt 120ttatttgttt gttgctg atg ggt ctt ggg ttc aag atg cag cag age cac 170ttcaactttc aaaacaaagt ggatgggatc ttcatcttat ataaccacac atcaatcatt 60tgtgctactt ctccaatttt ctttagagat gaaatgaaga gagtaagcaga caagacaagt 120ttatttgttt gttgctg atg ggt ctt gg cgag

Met Gly Leu Gly Phe Lys Met Gln Gln Ser His1 5 10Met Gly Read Gly Phe Lys Met Gln Gln Being His1 5 10

cac cct gtg gct ttg aag tta cat gag caa gct ggg age cag aga aag 218His Pro vai Ala Leu Lys Leu His Glu Gln Ala Gly Ser Gln Arg Lys15 20 25cac cct gtg gtt ttg aag tta cat gag caa gct ggg age cag aga aag 218His Pro Goes Wing Leu Lys Leu His Glu Gln Wing Gly Ser Gln Arg Lys15 20 25

ttc act ttc att cag aac ttc aga aac tgg ttt cta ccc ctt ctg ttt 266Phettc act ttc att cag aac ttc aga aac tgg ttt cta ccc ctt ctg ttt 266Phe

gtavaigtavai

atgMet60tgtCysatgMet60tgtCys

aacAsnaacAsn

agaArgagaArg

aggArgaggArg

agaArg140gttvaiagaArg140gttvai

gcaAlagcaAla

gatgat

ASpASp

2020

2525

Thr Phe30tgg ttcTrp Phe45Thr Phe30tgg ttcTrp Phe45

gat gctAsp Alagat gctAsp Wing

gat caaAsp Glngat caaAsp Gln

cat gtcHis vaicat gtcHis will

aat actAsn Thr110aca gcaThr Ala125aat actAsn Thr110aca gcaThr Ala125

gtg gttvai vaigtg gttvai will

ata aagIle LysaagIle Lys

gca gtgAla vaigca gtgAla go

atg atgMet Met190atg atgMet Met190

lie Gln Asn Phelie Gln Asn Phe

att gttIle vaiatt gttIle will

gaa actGlu ThrgaG actGlu Thr

agg gctArg Ala80cat gccHis Ala95agg gctArg Ala80cat gccHis Ala95

tca gccSer Alatca gccSer Wing

ttt gaaPhe Gluttt gaaPhe Glu

cac tcaHis Sercac tcaHis Ser

aca atgThr Met160ata tttIle Phe175aca atgThr Met160ata tttIle Phe175

tct gggSer Glytct gggSer Gly

atg gctMet Alaatg gctMet Wing

50aaa gtcLys vai6550aaa gtcLys vai65

aga atgArg Metaga atgArg Met

ctt gccLeu Alactt gccLeu Ala

att gacIle Aspatt gacIle Asp

cgg ccaArg Pro130gag agaGlu Arg145cgg ccaArg Pro130gag agaGlu Arg145

gaa agaGlu Arggaa agaGlu Arg

Arg Asn35Arg Asn35

gca tttAla Phegca tttAla Phe

aga aggArg Argaga aggArg Arg

cta caaLeu Glncta caaLeu Gln

ate cttlie Leu100cag gaaGln Glu115until cttlie Leu100cag gaaGln Glu115

tta atgLeu Mettta atgLeu Met

gaa agaGlu Arggaa agaGlu Arg

gtg cctvai Progtg cctvai Pro

Trp Phe Leu Pro Leu Leu Phe40Trp Phe Leu Pro Leu Leu Phe40

ate tac cat aaaIle Tyr His LysTil his cat Tyr His Lys

gca cag gaa actAla Gln Glu Thr180gca cag gaa actAla Gln Glu Thr180

gag gag gac cgaGlu Glu Asp Arg195gag gag gac cgaGlu Glu Asp Arg195

ggt gcc tgcGly Ala Cys55ggt gcc tgcGly Wing Cys55

aaa gag gtgLys Glu vai70aaa gag gtgLys Glu vai70

gac caa ttcAsp Gln Phe85gac caa ttcAsp Gln Phe85

gtt tca accVal Ser Thrgtt tca accVal Ser Thr

acc ttt gcaThr Phe Alaacc ttt gcaThr Phe Ala

agt ggg gtgSer Gly Val135agt ggg gtgSer Gly Val135

ttt gag aagPhe Glu Lysttt gag aagPhe Glu Lys

150tca ggg gttSer Gly Val165150tca ggg gttSer Gly Val165

gtc tct tacvai Ser Tyrgtc tct tacvai Ser Tyr

gag aac attGlu Asn Ilegag aac attGlu Asn Ile

314314

ctg ggtLeu Glyctg ggtLeu Gly

agt gtcSer Valagt gtcSer Val

ttc catPhe His105gaa tacGlu Tyr120ttc catPhe His105gaa tacGlu Tyr120

gcc tatAla Tyrgcc tatAla Tyr

caa catGln HiscatGln His

agg gatArg Aspagg gatArg Asp

ctt gaaLeu Glu185ttg aggLeu Arg200ctt gaaLeu Glu185ttg aggLeu Arg200

age ctcSer Leu75agt gtcser vai90age ctcSer Leu75agt gtcser vai90

tac tacTyr Tyrtac tacTyr Tyr

acg gccThr Alaacg gccThr Wing

gca cagAla Glngca cagAla Gln

ggg tggggg tgg

Gly Trp155gag tatGlu Tyr170Gly Trp155gag tatGlu Tyr170

tct attSer Iletct attSer Ile

gct agaAla Arggct agaAla Arg

362362

410410

458458

506506

554554

602602

650650

698698

746gcc act ggg aaa gct gtt ctg act age cct ttc aga ctg ctg gat tet 794746gcc act ggg aaa gct gtt ctg act age cct ttc aga ctg ctg gat tet 794

Ala Thr Gly Lys Ala vai Leu Thr Ser Pro Phe Arg Leu Leu Asp SerWing Thr Gly Lys Wing Go Read Thr Be Pro Phe Arg Read Le Asp Ser

205 210 215205 210 215

cat cat ctt ggc gtg gtt cta aca ttt cct gtt tat aaa tet aag ctc 842His His Leu Gly vai Val Leu Thr Phe Pro vai Tyr Lys Ser Lys Leu220 225 230 235cat cat ctt ggc gtg gtt cta aca ttt cct gtt tat aaa tet aag ctc 842His His Leu Gly will Val Leu Thr Phe Pro will Tyr Lys Ser Lys Leu220 225 230 235

cct cca gag cca acg acg gaa gag gtc att aaa gcc ata gea gga tat 890Pro Pro Glu Pro Thr Thr Glu Glu vai Ile Lys Ala Ile Ala Gly Tyr240 245 250cct cca gag cca acg acg gaa gag gtc att aaa gcc ata gea gga tat 890Pro Pro Glu Pro Thr Thr Glu Glu will Ile Lys Ala Ile Ala Gly Tyr240 245 250

att gga gga tcc ttt gat gtt gag tcc ctt gtg gag aat tta ctt ggt 938Ile Gly Gly Ser Phe Asp vai Glu Ser Leu Val Glu Asn Leu Leu Glyatt gga gga tcc ttt gat gtt gag tcc ctt gtg gag aat tta ctt ggt 938Ile Gly Gly Ser Phe Asp will Glu Ser Leu Val Glu Asn Leu Leu Gly

255 260 265255 260 265

caa ctt gct ggt aac caa gea att ttg gtg aag gta tat gat ata aca 986Gln Leu Ala Gly Asn Gln Ala Ile Leu Val Lys Val Tyr Asp Ile Thr 270 275 280caa ctt gct ggt aac caa gea att ttg gtg aag gta tat gat ata aca 986Gln Leu Wing Gly Asn Gln Wing Ile Leu Val Lys Val Tyr Asp Ile Thr 270 275 280

aac tet age gac ccc cta ate atg tat ggc age caa tat gaa gag ggt 1034Asn Ser Ser Asp Pro Leu Ile Met Tyr Gly Ser Gln Tyr Glu Glu Glyaac tet age gac ccc cta till atg tat ggc age caa tat gaa gag ggt 1034Asn Ser Ser Asp Pro Leu Ile Met Tyr Gly Ser Gln Tyr Glu Glu Gly

285 290 295285 290 295

gat atg tet ctt gtc cat gaa agt aag ctt gat ttt gga gat cca tac 1082Asp Met Ser Leu vai His Glu Ser Lys Leu Asp Phe Gly Asp Pro Tyr300 305 310 315gat atg tet ct gtc cat gaa agt aag ctt gat ttt gga gat cca tac 1082Asp Met Ser Leu Goes His Glu Ser Lys Leu Asp Phe Gly Asp Pro Tyr300 305 310 315

agg aaa cat cac atg ate tgt aga tat cac caa cag gea cca aca aat 1130Arg Lys His His Met Ile Cys Arg Tyr His Gln Gln Ala Pro Thr Asn320 325 330agg aaa cat cac atg till tgt aga tat cac caa cag gea cca aca aat 1130Arg Lys His His Met Ile Cys Arg Tyr His Gln Ala Pro Thr Asn320 325 330

tgg ata gea tat acc acg gea ttc cta ttc ttt gtg att ctt tgt tta 1178Trp Ile Ala Tyr Thr Thr Ala Phe Leu Phe Phe Val Ile Leu Cys Leutgg ata gea tat acc acg gea ttc cta ttc ttt gtg att ctt tgt tta 1178Trp Ile Wing Tyr Thr Thr Wing Phe Leu Phe Val Ile Leu Cys Leu

335 340 345335 340 345

gtg ggt tac att tta tat gct gct gga act cac att gtc aag gta gaa 1226Val Gly Tyr Ile Leu Tyr Ala Ala Gly Thr His Ile Val Lys Val Glu 350 355 360gtg ggt tac att tta tat gct gct gga act cac att gtc aag gta gaa 1226Val Gly Tyr Ile Leu Tyr Wing Gly Thr His Ile Val Lys Val Glu 350 355 360

gat gat tac aat gea atg cag gat tta aaa gtc aaa gea gaa gea gct 1274Asp Asp Tyr Asn Ala Met Gln Asp Leu Lys Val Lys Ala Glu Ala Ala365 370 375gat gat tac aat gea atg cag gat tta aaa gtc aaa gea gaa gea gct 1274Asp Asp Tyr Asn Ala Met Gln Asp Leu Lys Val Lys Ala Glu Ala Ala365 370 375

gat att gcc aag tca cag ttt cta gct acc gtc tct cat gaa att aga 1322Asp Ile Ala Lys Ser Gln Phe Leu Ala Thr vai Ser His Glu Ile Arg380 385 390 395gat att gcc aag tca cag ttt cta gct acc gtc tct cat gaa att aga 1322Asp Ile Wing Lys Ser Gln Phe Wing Wing Thr Will Be His Glu Ile Arg380 385 390 395

act ccc atg aat gga att tta gga atg ctt ggt ctg ctt tta cgc aca 1370Thr Pro Met Asn Gly Ile Leu Gly Met Leu Gly Leu Leu Leu Arg Thract ccc atg aat gga att tta gga atg ctt ggt ctg ctt tta cgc aca 1370Thr Pro Met Asn Gly Ile Leu Gly Met Leu Gly Leu Leu Arg Thr

400 405 410400 405 410

gaa ttg agt tca aca caa aga gac tat gct cag act gct caa gca tgt 1418Glu Leu Ser Ser Thr Gln Arg Asp Tyr Ala Gln Thr Ala Gln Ala Cys 415 420 425gaa ttg agt tca aca caa aga gac tat gct cag act gct caa gca tgt 1418Glu Leu Ser Ser Thr Gln Arg Asp Tyr Ala Gln Thr Ala Gln Ala Cys 415 420 425

ggg aag gca cta ata gca tta ata aat gag gtg ctt gac cga gct aaa 1466Gly Lys Ala Leu Ile Ala Leu Ile Asn Glu Val Leu Asp Arg Ala Lysggg aag gca cta ata gca tta ata aat gag gtg ctt gac cga gct aaa 1466Gly Lys Ala Leu Ile Ala Leu Ile Asn Glu Val Leu Asp Arg Ala Lys

430 435 440430 435 440

att gaa gca ggc aaa tta gag cta gaa gca gtt cca ttt gac ctt cgt 1514 lie Glu Ala Gly Lys Leu Glu Leu Glu Ala Val Pro Phe Asp Leu Arg445 450 455att gaa gca ggc aaa tta gag cta gaa gca gtt cca ttt gac ctt cgt 1514 lie Glu Ala Gly Lys Leu Glu Leu Glu Ala Val Pro Phe Asp Leu Arg445 450 455

tcc ata ctt gac gat gtc ctt tct ctt ttt tct gag aag tca aga cac 1562Ser lie Leu Asp Asp Val Leu Ser Leu Phe Ser Glu Lys Ser Arg His460 465 470 475tcc ata ctt gac gat gtc ctt tct ctt ttt tct gag aag tca aga cac 1562See lie Leu Asp Asp Val Leu Ser Leu Phe Ser Glu Lys Ser Arg His460 465 470 475

aaa ggc tta gag ctg gca gtg ttt gtt tct gac aaa gtt ccg gat ata 1610Lys Gly Leu Glu Leu Ala vai Phe vai Ser Asp Lys Val Pro Asp Ileaaa ggc tta gag ctg gca gtg ttt gtt tct gac aaa gtt ccg gat ata 1610Lys Gly Leu Glu Leu Ala Will Phe Will Be Asp Lys Val Pro Asp Ile

480 485 490480 485 490

gtt atg ggc gat cct ggg cga ttc aga caa ata gtg aca aat ctt gtt 1658Val Met Gly Asp Pro Gly Arg Phe Arg Gln Ile vai Thr Asn Leu Val 495 500 505gt atg ggc gat cct ggg cga ttc aga caa ata gtg aca aat ctt gtt 1658Val Met Gly Asp Pro Gly Arg Phe Arg Gln Ile Go Thr Asn Leu Val 495 500 505

gga aac tct gtt aag ttc act gag cga ggt cat ata ttt gtt aaa gtc 1706Gly Asn Ser Val Lys Phe Thr Glu Arg Gly His Ile Phe vai Lys Valgga aac tct gtt aag ttc act gag cga ggt cat ata ttt gtt aaa gtc 1706Gly Asn Ser Val Lys Phe Thr Glu Arg Gly His Ile Phe Goes Lys Val

510 515 520510 515 520

cat tta gct gaa aaa aga cag tgc aca atg aat gga aaa tgt gag act 175430 His Leu Ala Glu Lys Arg Gln Cys Thr Met Asn Gly Lys Cys Glu Thr525 530 535cat tta gct gaa aaa aga cag tgc aca atg aat gga aaa tgt gag act 175430 His Leu Wing Glu Lys Arg Gln Cys Thr Met Asn Gly Lys Cys Glu Thr525 530 535

ttt cta aat gga ggc tgt gat gat gtt ttg cat gta tct ggc agt tat 1802Phe Leu Asn Gly Gly Cys Asp Asp Val Leu His Val Ser Gly Ser Tyr540 545 550 555ttt cta aat gga ggc tgt gat gat gtt ttg cat gta tct ggc agt tat 1802Phe Leu Asn Gly Gly Cys Asp Asp Val Leu His Val Ser Gly Ser Tyr540 545 550 555

aat ttg aaa acc ctt agt gga tat gaa gcc gct gat gaa cgg aac age 1850Asn Leu Lys Thr Leu Ser Gly Tyr Glu Ala Ala Asp Glu Arg Asn Ser 560 565 570aat ttg aaa acc ctt agt gga tat gaa gcc gct gat gaa cgg aac age 1850Asn Leu Lys Thr Leu Ser Gly Tyr Glu Ala Wing Asp Glu Arg Asn Ser 560 565 570

tgg gat aat ttt aag cat cat att gct gac gaa gaa ttt ttc ttt gat 1898Trp Asp Asn Phe Lys His His Ile Ala Asp Glu Glu Phe Phe Phe Asptgg gat aat ttt aag cat cat att gct gac gaa gaa ttt ttc ttt gat 1898Trp Asp Asn Phe Lys His His Ile Ala Asp Glu Glu Phe Phe Phe Asp

575 580 585575 580 585

gct tcg gtt aaa aag ttg gcc tet agt gaa tet tat gag caa gtc acc 1946Ala Ser vai Lys Lys Leu Ala Ser Ser Glu Ser Tyr Glu Gln Val Thr590 595 600gct tcg gtt aaa aag ttg gcc tet agt gaa tet tat gag caa gtc acc 1946Ala Ser will Lys Lys Leu Ala Ser Be Glu Ser Tyr Glu Gln Val Thr590 595 600

ttg atg gtc age gtg gag gac act gga att ggg att tet ttc tet gcc 1994Leu Met Val Ser Val Glu Asp Thr Gly Ile Gly Ile Ser Phe Ser Ala605 610 615ttg atg gtc age gtg gag gac act gga att ggg att tet ttc tet gcc 1994Leu Met Val Ser Val Glu Asp Thr Gly Ile Gly Ile Ser Phe Ser Ala605 610 615

caa gat agt att ttc atg cct ttt gtg cag gct gac age tca acc tet 2042Gln Asp Ser Ile Phe Met Pro Phe vai Gln Ala Asp ser Ser Thr Ser620 625 630 635caa gat agt att ttc atg cct ttt gtg cag gct gac age tca acc tet 2042Gln Asp Ser Ile Phe Met Pro Phe Go Gln Ala Asp Be Ser Thr Ser620 625 630 635

cga aac tat ggg ggt acc ggg ate ggc ttg agt ate agt aag tgc ttg 2090Arg Asn Tyr Gly Gly Thr Gly Ile Gly Leu ser Ile Ser Lys Cys Leu640 645 650cga aac tat ggg ggt acc ggg until ggc ttg agt until agt aag tgc ttg 2090Arg Asn Tyr Gly Gly Thr Gly Ile Gly Leu be Ile Ser Lys Cys Leu640 645 650

gtt gaa ctg atg ggc ggt cag ata aac ttc ata age cga ccc cag gtc 2138vai Glu Leu Met Gly Gly Gln Ile Asn Phe Ile ser Arg Pro Gln Valgtt gaa ctg atg ggc ggt cag ata aac ttc ata age cga ccc cag gtc 2138vai Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Be Arg Pro Gln Val

655 660 665655 660 665

ggg age acg ttt tca ttc act gea gat ttc gga aca ttt aag aaa aac 2186Gly Ser Thr Phe Ser Phe Thr Ala Asp Phe Gly Thr Phe Lys Lys Asn670 675 680ggg age acg ttt tca ttc act gea gat ttc gga aca ttt aag aaa aac 2186Gly Be Thr Phe Be Phe Thr Wing Asp Phe Gly Thr Phe Lys Lys Asn670 675 680

tca aca act gac atg aag aaa ctt aac ttt gaa gat cta cct tet agt 2234Ser Thr Thr Asp Met Lys Lys Leu Asn Phe Glu Asp Leu Pro Ser Ser685 690 695tca aca act gac atg aag aaa ctt aac ttt gaa gat cta cct tet agt 2234Ser Thr Thr Asp Met Lys Lys Leu Asn Phe Glu Asp Leu Pro Ser685 690 695

ttt aga ggt ctt aaa gcc att gtg gtt gat gga aaa cct gtt aga gct 2282Phe Arg Gly Leu Lys Ala Ile vai vai Asp Gly Lys Pro Val Arg Ala700 705 710 715gca gtg act aga tac cat ttg aag aga cta ggg ata caa gct aaa gtt 2330Ala Val Thr Arg Tyr His Leu Lys Arg Leu Gly Ile Gln Ala Lys Valttt aga ggt ctt aaa gcc att gtg gtt gat gga aaa cct gtt aga gct 2282Phe Arg Gly Leu Lys Ala Ile Goes Go Asp Gly Lys Pro Val Arg Ala700 705 710 715gca gtg act aga tac cat ttg aag aga cta ggg ata caa gct 2330Ala Val Thr Arg Tyr His Leu Lys Arg Leu Gly Ile Gln Ala Lys Val

720 725 730720 725 730

gca att age ate aat aag gct gtt tet tta tgt ggg aaa aat ggt tet 2378Ala lie ser Ile Asn Lys Ala Val ser Leu Cys Gly Lys Asn Gly Ser735 740 745gca att age until aat aag gct gtt tet tta tgt ggg aaa aat ggt tet 2378Ala lie be Ile Asn Lys Ala Val be Leu Cys Gly Lys Asn Gly Ser735 740 745

ttg acc tcg gca tta ttt cag cct gat att att ttt gtt gag aag gac 2426Leu Thr Ser Ala Leu Phe Gln Pro Asp Ile lie Phe Val Glu Lys Asp750 755 760ttg acc tcg gca tta ttt cag cct gat att att ttt gtt gag aag gac 2426Leu Thr Ser Wing Leu Phe Gln Pro Asp Ile lie Phe Val Glu Lys Asp750 755 760

tet tgg gtt tet gga gag gat ggt ggt ate ttc aat gcg ttt aag atg 2474Ser Trp Val Ser Gly Glu Asp Gly Gly Ile Phe Asn Ala Phe Lys Mettet tgg gtt tet gga gag gat ggt ggt until ttc aat gcg ttt aag atg 2474Ser Trp Val Ser Gly Glu Asp Gly Ile Phe Asn Ala Phe Lys Met

765 770 775765 770 775

cct caa atg ate ctt ctt gca acc aat ate tgt aac gct gaa ttt gat 2522Pro Gln Met Ile Leu Leu Ala Thr Asn Ile Cys Asn Ala Glu Phe Asp 780 785 790 795cct caa atg till ctt ctt gca acc aat till tgt aac gct gaa ttt gat 2522Pro Gln Met Ile Leu Read Ala Thr Asn Ile Cys Asn Ala Glu Phe Asp 780 785 790 795

aaa gcc aaa gct gca ggt ttc agt gat aca gtg ate atg aag cca ctg 2570Lys Ala Lys Ala Ala Gly Phe Ser Asp Thr vai Ile Met Lys Pro Leuaaa gcc aaa gct gca ggt ttc agt gat aca gtg till atg aag cca ctg 2570Lys Ala Lys Ala Ala Gly Phe Ser Asp Thr Go Ile Met Lys Pro Leu

800 805 810800 805 810

aga gct agt atg ctg gct gct tgt ctt cag caa gtt ttc ggg act ggc 2618Arg Ala Ser Met Leu Ala Ala Cys Leu Gln Gln vai Phe Gly Thr Gly815 820 825aga gct agt atg ctg gct gct tgt ctt cag caa gtt ttc ggg act ggc 2618Arg Wing Ser Met Leu Wing Cys Wing Gln Gln Go Phe Gly Thr Gly815 820 825

aag acg agg cag ttt ggg aaa gac atg tcg aat ggt tet tca gta cga 2666Lys Thr Arg Gln Phe Gly Lys Asp Met Ser Asn Gly Ser Ser Val Arg830 835 840aag acg agg cag ttt ggg aaa gac atg tcg aat ggt tet tca gta cga 2666Lys Thr Arg Gln Phe Gly Lys Asp Met Ser Asn Gly Ser Ser Val Arg830 835 840

age ctt ctt tgc gga aag aaa ate tta gtg gtt gat gat aat ttg gtg 2714Ser Leu Leu Cys Gly Lys Lys Ile Leu Val Val Asp Asp Asn Leu vaiage ctt ctt tgc gga aag aaa till tta gtg gtt gat gat aat ttg gtg 2714Ser Leu Leu Cys Gly Lys Lys Ile Leu Val Val Asp Asp Asn Leu Goes

845 850 855845 850 855

aac cga agg gtc gcc gcc ggc gcg ttg aaa aac ttt gga gct gat gtc 2762Asn Arg Arg vai Ala Ala Gly Ala Leu Lys Asn Phe Gly Ala Asp Val860 865 870 875aac cga agg gtc gcc gcc ggc gcg ttg aaa aac ttt gga gct gat gtc 2762Asn Arg Arg goes Wing Gly Wing Wing Leu Lys Asn Phe Gly Wing Asp Val860 865 870 875

aaa tgt gca gca agt ggc aaa gct gct ctt gaa atg ctt caa tat cct 2810Lys Cys Ala Ala Ser Gly Lys Ala Ala Leu Glu Met Leu Gln Tyr Pro880 885 890aaa tgt gca gca agt ggc aaa gct gct ctt gaa atg ctt caa tat cct 2810Lys Cys Wing Ala Ser Gly Lys Wing Wing Leu Glu Met Leu Gln Tyr Pro880 885 890

cac gat ttc gat gct tgc ttc atg gat att caa atg cca gaa atg gat 2858His Asp Phe Asp Ala Cys Phe Met Asp Ile Gln Met Pro Glu Met Asp895 900 905cac gat ttc gat gct tgc ttc atg gat att caa atg cca gaa atg gat 2858His Asp Phe Asp Cys Phe Met Asp Ile Gln Met Pro Glu Met Asp895 900 905

ggg ttt gag gcc act cgt cga att cgg atg atg gaa aga gag gca agt 2906Gly Phe Glu Ala Thr Arg Arg Ile Arg Met Met Glu Arg Glu Ala Serggg ttt gag gcc act cgt cga att cgg atg atg gaa aga gag gca agt 2906Gly Phe Glu Ala Thr Arg Arg Ile Arg Met Met Glu Arg Glu Ala Ser

910 915 920910 915 920

gag cag ctg aaa agt gaa tct ggt gaa gaa aat ggt aag aaa agt gag 2954Glu Gln Leu Lys Ser Glu Ser Gly Glu Glu Asn Gly Lys Lys Ser Glu925 9B0 935gag cag ctg aaa agt gaa tct ggt gaa gaa aat ggt aag aaa agt gag 2954Glu Gln Leu Lys Be Glu Ser Gly Glu Asn Gly Lys Lys Ser Glu925 9B0 935

ttc cac atg cct ata ttg gcc atg aca gct gat gta ate cat gct aca 3002Phe His Met Pro Ile Leu Ala Met Thr Ala Asp vai Ile His Ala Thr940 945 950 955ttc cac atg cct ata ttg gcc atg aca gct gat gta up to cat gct aca 3002Phe His Met Pro Ile Read Ala Met Thr Ala Asp will Ile His Ala Thr940 945 950 955

tat gat aag tgc tta aat tgt ggg atg gat gga tac gtc tca aag cct 3050Tyr Asp Lys Cys Leu Asn Cys Gly Met Asp Gly Tyr vai Ser Lys Protat gat aag tgc tta aat tgt ggg atg gat gga tac gtc tca aag cct 3050Tyr Asp Lys Cys Leu Asn Cys Gly Met Asp Gly Tyr Will Be Lys Pro

960 965 970960 965 970

ttt gaa gaa gag aat ctc tat caa gca gtt gca aag ttc ttc aag tcc 3098Phe Glu Glu Glu Asn Leu Tyr Gln Ala vai Ala Lys Phe Phe Lys Ser975 980 985ttt gaa gaa gag aat ctc tat caa gca gtt gca aag ttc ttc aag tcc 3098Phe Glu Glu Glu Asn Leu Tyr Gln Wing Go Ala Lys Phe Phe Lys Ser975 980 985

aag cct gct tca gac tca tga cactgcttat tctgcagaac aggtcaacca 3149aag cct gct tca gac tca tga cactgcttat tctgcagaac aggtcaacca 3149

Lys Pro Ala Ser Asp Ser990Lys Pro Wing Ser Asp Ser990

acttttgatt gagaaacatt tagtgttagc atgtttggat caacttctcc cagcatcaat 3209tctgaaactc agaagctact catagaagct tctagccaga attgattttg gcttcagaag 3269ctactaatgg ttttatgtag agagcaaaag ttggtttcca agatatgcga ggatccatga 3329tgatacacac gtctgaagtg ateattteta accagttgaa gtttcactcg acgtgatttg 3389aatccaagta aatgcatacc acataattat ccatccccat cttgtgtaca gattctccca 3449agggataaga aaatttatgt aaattcaatt tttttctttt gcatctcaat acttccctgt 3509tagaactttt tccctatgat tatttccact tttcattttc aattatattt tttgtaaaa 3568acttttgatt gagaaacatt tagtgttagc atgtttggat caacttctcc cagcatcaat 3209tctgaaactc agaagctact catagaagct tctagccaga attgattttg gcttcagaag 3269ctactaatgg ttttatgtag agagcaaaag ttggtttcca agatatgcga ggatccatga 3329tgatacacac gtctgaagtg ateattteta accagttgaa gtttcactcg acgtgatttg 3389aatccaagta aatgcatacc acataattat ccatccccat cttgtgtaca gattctccca 3449agggataaga aaatttatgt aaattcaatt tttttctttt gcatctcaat acttccctgt 3509tagaactttt tccctatgat tatttccact tttcattttc aattatattt 3568 tttgtaaaa

<210> 3<211> 993<212> PRT<213> Lotus japonicus<400> 3<210> 3 <211> 993 <212> PRT <213> Lotus japonicus <400> 3

Met Gly Leu Gly Phe Lys Met GTn Gln Ser His His Pro Val Ala Leu15 10 15Met Gly Leu Gly Phe Lys Met GTn Gln To Be His His Pro Val Wing Leu15 10 15

Lys Leu His Glu Gln Ala Gly Ser Gln Arg Lys Phe Thr Phe Ile Gln20 25 30Lys Read His Glu Gln Wing Gly Be Gln Arg Lys Phe Thr Phe Ile Gln20 25 30

Asn Phe Arg Asn Trp Phe Leu Pro Leu Leu Phe Val Trp Phe Ile ValAsn Phe Arg Asn Trp Phe Leu Pro Leu Leu Phe Val Trp Phe Ile Val

35 40 4535 40 45

Met Ala Ala Phe Gly Ala Cys Ile Tyr His Lys Met Asp Ala Glu Thr 50 55 60Met Wing Phe Wing Gly Cys Wing Ile Tyr His Lys Met Asp Wing Glu Thr 50 55 60

Lys Val Arg Arg Lys Glu Val Leu Gly Ser Leu Cys Asp Gln Arg Ala65 70 75 80Lys Val Arg Arg Lys Glu Val Leu Gly Ser Leu Cys Asp Gln Arg Ala65 70 75 80

Arg Met Leu Gln Asp Gln Phe Ser Val Ser Val Asn His vai His Ala85 90 95Arg Met Read Gln Asp Gln Phe Ser Val Ser Val Asn His Goes His Ala85 90 95

Leu Ala Ile Leu Val Ser Thr Phe His Tyr Tyr Arg Asn Thr Ser Ala100 105 110Leu Ala Ile Leu Val Be Thr Phe His Tyr Tyr Arg Asn Thr Be Ala100 105 110

Ile Asp Gln Glu Thr Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe GluIle Asp Gln Glu Thr Phe Wing Glu Tyr Glou Arg Wing Thr Phe Glu

115 120 125115 120 125

Arg Pro Leu Met Ser Gly Val Ala Tyr Ala Gln Arg Val vai His Ser 130 135 140Arg Pro Read Met Ser Gly Val Wing Tyr Wing Gln Arg Val Goes His Ser 130 135 140

Glu Arg Glu Arg Phe Glu Lys Gln His Gly Trp vai Ile Lys Thr Met145 150 155 160Glu Arg Glu Arg Phe Glu Lys Gln His Gly Trp Goes Ile Lys Thr Met145 150 155 160

Glu Arg Val Pro Ser Gly Val Arg Asp Glu Tyr Ala Ala Val Ile Phe165 170 175Glu Arg Val Pro Ser Gly Val Arg Asp Glu Tyr Wing Val Ile Wing Phe165 170 175

Ala Gln Glu Thr Val Ser Tyr Leu Glu Ser Ile Asp Met Met Ser Gly180 185 190Gln Wing Glu Thr Val Ser Tyr Leu Glu Ser Ile Asp Met Met Ser Gly180 185 190

Glu Glu Asp Arg Glu Asn Ile Leu Arg Ala Arg Ala Thr Gly Lys AlaGlu Glu Asp Arg Glu Asn Ile Read Leu Arg Wing Arg Wing Thr Gly Lys Wing

195 200 205195 200 205

Val Leu Thr ser Pro Phe Arg Leu Leu Asp Ser His His Leu Gly Val 210 215 220Val Leu Thr Be Pro Phe Arg Leu Read Asp Be His His Leu Gly Val 210 215 220

Val Leu Thr Phe Pro Val Tyr Lys Ser Lys Leu Pro Pro Glu Pro Thr225 230 235 240Thr Glu Glu vai Ile Lys Ala Ile Ala Gly Tyr Ile Gly Gly Ser PheVal Leu Thr Phe Pro Val Tyr Lys Ser Lys Leu Pro Pro Glu Pro Thr225 230 235 240Thr Glu Glu Go Ile Lys Wing Ile Wing Gly Tyr Ile Gly Gly Ser Phe

245 250 255245 250 255

Asp vai Glu Ser Leu vai Glu Asn Leu Leu Gly Gln Leu Ala Gly Asn260 265 270Asp goes Glu Be Leu goes Glu Asn Leu Leu Gly Gln Leu Wing Gly Asn260 265 270

Gln Ala Ile Leu vai Lys Val Tyr Asp Ile Thr Asn ser Ser Asp Pro275 280 285Gln Wing Ile Leu Will Lys Val Tyr Asp Ile Thr Asn Be Ser Asp Pro275 280 285

Leu Ile Met Tyr Gly Ser Gln Tyr Glu Glu Gly Asp Met Ser Leu ValLeu Ile Met Tyr Gly Ser Gln Tyr Glu Glu Gly Asp Met Ser Leu Val

290 295 300290 295 300

His Glu Ser Lys Leu Asp Phe Gly Asp Pro Tyr Arg Lys His His Met 305 310 315 320His Glu Ser Lys Read Asp Phe Gly Asp Pro Tyr Arg Lys His His 305 310 315 320

Ile Cys Arg Tyr His Gln Gln Ala Pro Thr Asn Trp Ile Ala Tyr ThrIle Cys Arg Tyr His Gln Gln Wing Pro Thr Asn Trp Ile Cys Wing Tyr Thr

325 330 335325 330 335

Thr Ala Phe Leu Phe Phe Val Ile Leu Cys Leu Val Gly Tyr Ile Leu340 345 350Thr Wing Phe Leu Phe Phe Val Ile Leu Cys Leu Val Gly Tyr Ile Leu340 345 350

Tyr Ala Ala Gly Thr His Ile vai Lys vai Glu Asp Asp Tyr Asn Ala355 360 365Tyr Wing Ally Gly Thr His Ile Goes Lys Goes Glu Asp Asp Tyr Asn Ala355 360 365

Met Gln Asp Leu Lys Val Lys Ala Glu Ala Ala Asp Ile Ala Lys SerMet Gln Asp Leu Lys Val Lys Wing Glu Wing Asp Wing Ile Wing Lys Ser

370 375 380370 375 380

Gln Phe Leu Ala Thr Val Ser His Glu Ile Arg Thr Pro Met Asn Gly1 - 20 385 390 395 400Gln Phe Read Wing Thr Val Be His Glu Ile Arg Thr Pro Met Asn Gly1 - 20 385 390 395 400

Ile Leu Gly Met Leu Gly Leu Leu Leu Arg Thr Glu Leu Ser Ser ThrIle Leu Gly Met Leu Gly Leu Leu Leu Arg Thr Glu Leu Ser Be Thr

405 410 415405 410 415

Gln Arg Asp Tyr Ala Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu Ile420 425 430Gln Arg Asp Tyr Wing Gln Thr Wing Gln Cys Wing Gly Lys Wing Leu Ile420 425 430

Ala Leu Ile Asn Glu Val Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys435 440 445Wing Leu Ile Asn Glu Val Leu Asp Arg Wing Lys Ile Glu Wing Gly Lys435 440 445

Leu Glu Leu Glu Ala vai Pro Phe Asp Leu Arg Ser Ile Leu Asp AspLeu Glu Leu Glu Wing goes to Phe Asp Leu Arg Ser Ile Leu Asp Asp

450 455 460450 455 460

vai Leu Ser Leu Phe Ser Glu Lys Ser Arg His Lys Gly Leu Glu Leu 465 470 475 480will Read Be Read Be Phe Be Glu Lys Be Arg His Lys Gly Leu Glu Leu 465 470 475 480

Ala Val Phe Val ser Asp Lys vai Pro Asp Ile Val Met Gly Asp Pro485 490 495Gly Arg Phe Arg Gln lie vai Thr Asn Leu vai Gly Asn Ser vai LysWing Val Phe Val Be Asp Lys Go Pro Asp Ile Val Met Gly Asp Pro485 490 495Gly Arg Phe Arg Gln Lie Go Thr Asn Leu Go Gly Asn Be Go Lys

500 505 510500 505 510

Phe Thr Glu Arg Gly His Ile Phe vai Lys Val His Leu Ala Glu Lys515 520 525Phe Thr Glu Arg Gly His Ile Phe Go Lys Val His Leu Wing Glu Lys515 520 525

Arg Gln Cys Thr Met Asn Gly Lys Cys Glu Thr Phe Leu Asn Gly Gly530 535 540Arg Gln Cys Met Thr Asn Gly Lys Cys Glu Thr Phe Read Asn Gly Gly530 535 540

Cys Asp Asp Val Leu His vai Ser Gly Ser Tyr Asn Leu Lys Thr Leu545 550 555 560Cys Asp Asp Val Leu His Will Be Gly Ser Tyr Asn Leu Lys Thr Leu545 550 555 560

Ser Gly Tyr Glu Ala Ala Asp Glu Arg Asn ser Trp Asp Asn Phe Lys 565 570 575Be Gly Tyr Glu Wing Wing Asp Glu Arg Asn Be Trp Asp Asn Phe Lys 565 570 575

His His Ile Ala Asp Glu Glu Phe Phe Phe Asp Ala Ser Val Lys LysHis His Ile Wing Asp Glu Glu Phe Phe Phe Asp Wing Ser Val Lys Lys

580 585 590580 585 590

Leu Ala Ser Ser Glu Ser Tyr Glu Gln Val Thr Leu Met Val Ser vai595 600 605Leu Wing Be Ser Glu Ser Tyr Glu Gln Val Thr Read Leu Met Val Ser vai595 600 605

Glu Asp Thr Gly Ile Gly Ile Ser Phe Ser Ala Gln Asp Ser Ile Phe610 615 620Glu Asp Thr Gly Ile Gly Ile Be Phe Be Wing Gln Asp Be Ile Phe610 615 620

Met Pro Phe vai Gln Ala Asp Ser Ser Thr ser Arg Asn Tyr Gly Gly625 630 635 640Met Pro Phe Will Gln Wing Asp Be Be Thr Be Arg Asn Tyr Gly Gly625 630 635 640

Thr Gly lie Gly Leu Ser Ile Ser Lys Cys Leu vai Glu Leu Met Gly 645 650 655Thr Gly lie Gly Leu Be Ile Be Lys Cys Leu Goes Glu Leu Met Gly 645 650 655

Gly Gln lie Asn Phe Ile Ser Arg Pro Gln Val Gly Ser Thr Phe SerGly Gln lie Asn Phe Ile Be Arg Pro Gln Val Gly Be Thr Phe Be

660 665 670660 665 670

Phe Thr Ala Asp Phe Gly Thr Phe Lys Lys Asn Ser Thr Thr Asp Met675 680 685Phe Thr Wing Asp Phe Gly Thr Phe Lys Lys Asn Be Thr Thr Asp Met675 680 685

Lys Lys Leu Asn Phe Glu Asp Leu Pro Ser ser Phe Arg Gly Leu Lys690 695 700Lys Lys Leu Asn Phe Glu Asp Leu Pro To Be Phe Arg Gly Leu Lys690 695 700

Ala Ile vai Val Asp Gly Lys Pro vai Arg Ala Ala Val Thr Arg Tyr705 710 715 720Wing Ile goes Val Asp Gly Lys Pro goes Arg Wing Wing Val Thr Arg Tyr705 710 715 720

His Leu Lys Arg Leu Gly Ile Gln Ala Lys Val Ala Ile Ser Ile Asn 725 730 735His Leu Lys Arg Leu Gly Ile Gln Wing Lys Val Wing Ile Ser Ile Asn 725 730 735

Lys Ala Val Ser Leu Cys Gly Lys Asn Gly Ser Leu Thr Ser Ala Leu740 745 750Phe Gln Pro Asp Ile Ile Phe vai Glu Lys Asp Ser Trp vai ser GlyLys Wing Val Be Read Cys Gly Lys Asn Gly Be Read Leu Thr Be Wing Leu740 745 750Phe Gln Pro Asp Ile Ile Phe Will Glu Lys Asp Be Trp Will Be Gly

755 760 765755 760 765

Glu Asp Gly Gly Ile Phe Asn Ala Phe Lys Met Pro Gln Met Ile Leu770 775 780Glu Asp Gly Gly Ile Phe Asn Phe Lys Wing Pro Pro Gln Met Ile Leu770 775 780

Leu Ala Thr Asn Ile Cys Asn Ala Glu Phe Asp Lys Ala Lys Ala Ala785 790 795 800Leu Wing Thr Asn Ile Cys Asn Wing Glu Phe Asp Lys Wing Lys Wing Wing785 790 795 800

Gly Phe Ser Asp Thr vai Ile Met Lys Pro Leu Arg Ala Ser Met LeuGly Phe Ser Asp Thr Will Ile Met Lys Pro Read Arg Wing Be Met Met Leu

805 810 815805 810 815

Ala Ala Cys Leu Gln Gln Val Phe Gly Thr Gly Lys Thr Arg Gln Phe 820 825 830Wing Cys Wing Read Gln Gln Val Phe Gly Thr Gly Lys Thr Arg Gln Phe 820 825 830

Gly Lys Asp Met Ser Asn Gly Ser ser vai Arg Ser Leu Leu Cys GlyGly Lys Asp Met Be Asn Gly Be Be Go Arg Be Read Leu Cys Gly

835 840 845835 840 845

Lys Lys Ile Leu vai vai Asp Asp Asn Leu Val Asn Arg Arg Val Ala850 855 860Lys Lys Ile Leu Goes Go Asp Asp Asn Leu Val Asn Arg Arg Val Ala850 855 860

Ala Gly Ala Leu Lys Asn Phe Gly Ala Asp vai Lys Cys Ala Ala Ser865 870 875 880Wing Gly Wing Read Lys Asn Phe Gly Wing Asp Goes Lys Cys Wing Ser865 870 875 880

Gly Lys Ala Ala Leu Glu Met Leu Gln Tyr Pro His Asp Phe Asp AlaGly Lys Wing Wing Read Glu Met Read Gln Tyr Pro His Asp Phe Asp Wing

885 890 895885 890 895

Cys Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala Thr 900 905 910Cys Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Wing Thr 900 905 910

Arg Arg Ile Arg Met Met Glu Arg Glu Ala Ser Glu Gln Leu Lys SerArg Arg Ile Arg Met Met Glu Arg Glu Wing Be Glu Gln Leu Lys Ser

915 920 925915 920 925

Glu Ser Gly Glu Glu Asn Gly Lys Lys Ser Glu Phe His Met Pro Ile930 935 940Glu Being Gly Glu Glu Asn Gly Lys Lys Being Glu Phe His Met Pro Ile930 935 940

Leu Ala Met Thr Ala Asp Val Ile His Ala Thr Tyr Asp Lys Cys Leu945 950 955 960Leu Wing Met Thr Wing Asp Val Ile His Wing Thr Tyr Asp Lys Cys Leu945 950 955 960

Asn Cys Gly Met Asp Gly Tyr vai Ser Lys Pro Phe Glu Glu Glu AsnAsn Cys Gly Met Asp Gly Tyr Will Be Lys Pro Phe Glu Glu Glu Asn

965 970 975965 970 975

Leu Tyr Gln Ala Val Ala Lys Phe Phe Lys Ser Lys Pro Ala Ser Asp 980 985 990Read Tyr Gln Wing Val Wing Lys Phe Phe Lys Ser Lys Pro Asp Wing 980 985 990

Ser<210> 4<211> 12738<212> DNA<213> Lotus japonicus<220><221> promoter<222> (1)..(2359)<223> promoter + 5' UTR<220><221> exon<222> (2360)..(2704)<220><221> Intron<222> (2705)..(3024)<220><221> exon<222> (3025)..(3258)<220><221> Intron<222> (3259)..(3390)<220><221> exon<222> (3391)..(3560)<220><221> Intron<222> (3561)..(3834)<220><221> exon<222> (3835)..(4053)<223> C/T mutation in exon 4<220><221> Intron<222> (4054)..(4145)<220>Ser <210> 4 <211> 12738 <212> DNA <213> Lotus japonicus <220> <221> promoter <222> (1) .. (2359) <223> promoter + 5 'UTR <220> <221> exon <222> (2360) .. (2704) <220> <221> Intron <222> (2705) .. (3024) <220> <221> exon <222> (3025) .. (3258) <220 > <221> Intron <222> (3259) .. (3390) <220> <221> exon <222> (3391) .. (3560) <220> <221> Intron <222> (3561) .. ( 3834) <220> <221> exon <222> (3835) .. (4053) <223> C / T mutation in exon 4 <220> <221> Intron <222> (4054) .. (4145) <220 >

<221> exon<222> (4146)..(4332)<220><221> exon <222> (4146) .. (4332) <220>

<221> Intron<222> (4333)..(4420)<220><221> Intron <222> (4333) .. (4420) <220>

<221> exon<222> (4421)..(4472)<220><221> exon <222> (4421) .. (4472) <220>

<221> Intron<222> (4473)..(4586)<220><221> exon<221> Intron <222> (4473) .. (4586) <220> <221> exon

<222> (4587)..(4816)<220><222> (4587) .. (4816) <220>

<221> Intron<222> (4817)..(5037)<220><221> Intron <222> (4817) .. (5037) <220>

<221> exon<222> (5038)..(5136)<220><221> exon <222> (5038) .. (5136) <220>

<221> Intron<222> (5137)..(5308)<220><221> Intron <222> (5137) .. (5308) <220>

<221> exon<222> (5309)..(6024)<220><221> Intron<221> exon <222> (5309) .. (6024) <220> <221> Intron

<222> (6025)..(7042)<220><221> exon <222> (7043)..(7513) <220> <221> Intron <222> (7514)..(8306) <220> <221> exon <222> (8307)..(8564) <220> <221> Intron <222> (8565)..(12738) <22Β> 3' UTR + 3' intron <400> 4 cccgggattg ttcatatccc catgtgtgtg tttggtttgg acacaagcaa gcttgccaca 60gtggtgttta caactcattc tgaattatat atatatatat atatatataa ctaatctcta 120tcaataattt agttagtttt atgggataat atgattcccg tcaaattagc ttgtaggtta 180aaaatccata tttatataat aaattagact tatctaaatc acttaagtaa atatgatgta 240aaatcacaat agcttaggta gtttaacaat gttttatcat gaatttaaaa aaaattagtt 300gtttcaaaat attattggac caaataagca aagaggagct aatccaatta taaacagcac 360atctttataa ttgttttcta ctcgatctaa tattggttcc ttctttcctt gagaaaaaaa 420gggttaaaaa tgcgatttgt tttgtaaaat taatttgtga cattaacaag agaaaactct 480acttttttaa tttttaagaa gttacttaaa tatagaaaga aagaagttac ttaaatatga 540aacttgtcta accaaagtca accgatcaaa gcgtttggtt agcaaagaga acttaaaaaa 600ataactaaat taactgtttg aacagtgatg taacagagaa atagaatggc ttacatatgg 660attatggagt taatggaagg cacactagtt ttagttcacg gcaacaagac agaattatcc 720tcctatattt taggaggttt tcttatttat ttttaatgta agacgtgtat ccttcttttg 780aggtaataat gttcgagctt gaaacattca cattatagtg tgactaattg cttaatatga 840atcaagcatg taccacttga aacactcact catcgtttat aatataatta ctttatagta 900ctataaagta tgtaattatt ttctcctttt taagtatctc aatcattttt ttgacccaaa 960tattatttta gagaagaaac atgtcttaag accaatacat caacatatat ggagagtagt 1020ttgacaacct ctcatgagct ttctaaagcc aaacaaatgt actcacataa aaaaattaaa 1080aatttacagc aagtgagtta taaagagtga ttaataaatg ttgaataacg atggagggta 1140gttatgaaat tccataaata aagcaaagga tgtttatggc aattgacatg ggaataagat 1200<222> (6025) .. (7042) <220> <221> exon <222> (7043) .. (7513) <220> <221> Intron <222> (7514) .. (8306) <220> <221> exon <222> (8307) .. (8564) <220> <221> Intron <222> (8565) .. (12738) <22Β> 3 'UTR + 3' intron <400> 4 cccgggattg ttcatatccc catgtgtgtg tttggtttgg acacaagcaa gcttgccaca 60gtggtgttta caactcattc tgaattatat atatatatat atatatataa ctaatctcta 120tcaataattt agttagtttt atgggataat atgattcccg tcaaattagc ttgtaggtta 180aaaatccata tttatataat aaattagact tatctaaatc acttaagtaa atatgatgta 240aaatcacaat agcttaggta gtttaacaat gttttatcat gaatttaaaa aaaattagtt 300gtttcaaaat attattggac caaataagca aagaggagct aatccaatta taaacagcac 360atctttataa ttgttttcta ctcgatctaa tattggttcc ttctttcctt gagaaaaaaa 420gggttaaaaa tgcgatttgt tttgtaaaat taatttgtga cattaacaag agaaaactct 480acttttttaa tttttaagaa gttacttaaa tatagaaaga aagaagttac ttaaatatga 540aacttgtcta accaaagtca accgatcaaa gcgtttggtt agcaaagaga acttaaaaaa 600ataactaaat taactgtttg aacagtgatg ta acagagaa atagaatggc ttacatatgg 660attatggagt taatggaagg cacactagtt ttagttcacg gcaacaagac agaattatcc 720tcctatattt taggaggttt tcttatttat ttttaatgta agacgtgtat ccttcttttg 780aggtaataat gttcgagctt gaaacattca cattatagtg tgactaattg cttaatatga 840atcaagcatg taccacttga aacactcact catcgtttat aatataatta ctttatagta 900ctataaagta tgtaattatt ttctcctttt taagtatctc aatcattttt ttgacccaaa 960tattatttta gagaagaaac atgtcttaag accaatacat caacatatat ggagagtagt 1020ttgacaacct ctcatgagct ttctaaagcc aaacaaatgt actcacataa aaaaattaaa 1080aatttacagc aagtgagtta taaagagtga ttaataaatg ttgaataacg atggagggta 1140gttatgaaat tccataaata aagcaaagga tgtttatggc aattgacatg ggaataagat 1200

ccgcgcgttg tcgccgtgtc ataatcgctc agatttgtga tagcgagaga tttccattct 1260ccgcgcgttg tcgccgtgtc ataatcgctc agatttgtga tagcgagaga tttccattct 1260

tttcctcttt tttcgaacga acgaacgaac gaacacagta gcagctgtat taggattcag 1320tttcctcttt tttcgaacga acgaacgaac gaacacagta gcagctgtat taggattcag 1320

attgcatgat acattgatat tgatattgat attgatattg atattgttgt gggtcaagtc 1380attgcatgat acattgatat tgatattgat attgatattg atattgttgt gggtcaagtc 1380

tctactctac tatccaaagt aagcatatat atagagagag agagagagct tgggactagg 1440tctactctac tatccaaagt aagcatatat atagagagag agagagagct tgggactagg 1440

gaggatatta gcttatgtga ctgtgaagtt gaagggagac aagagcgtat ctggcaaaat 1500gaggatatta gcttatgtga ctgtgaagtt gaagggagac aagagcgtat ctggcaaaat 1500

cctcaaaata aaatactagt actagtacag agaaaagaga ctaagagaga gagtgctgct 1560cctcaaaata aaatactagt actagtacag agaaaagaga ctaagagaga gagtgctgct 1560

gcacatcaag acccattgtg atttgtgatt tgtgatttgt gatttgtgat ttgtgagtga 1620gcacatcaag acccattgtg atttgtgatt tgtgatttgt gatttgtgat ttgtgagtga 1620

gttcattgta caggtattat tgtttgttgt ttctctcctc aaccaccctc taaagtctaa 1680gttcattgta caggtattat tgtttgttgt ttctctcctc aaccaccctc taaagtctaa 1680

tctaactcat tgggctctgt gcttagctgg ttgtgttttg tgtatggtga attagggggt 1740tctaactcat tgggctctgt gcttagctgg ttgtgttttg tgtatggtga attagggggt 1740

caatctctgg ttttcatcat tattatatta tatatgggaa taccgtgctc tcttctttgt 1800caatctctgg ttttcatcat tattatatta tatatgggaa taccgtgctc tcttctttgt 1800

ccttcaccaa aactagtttc acgcctacac aacatgatta gagcctcttc atttttttaa 1860ccttcaccaa aactagtttc acgcctacac aacatgatta gagcctcttc atttttttaa 1860

tctcatcctt taagtgtatt tctatttcta ttggctattg ggacaagggg aaggtggtgc 1920tctcatcctt taagtgtatt tctatttcta ttggctattg ggacaagggg aaggtggtgc 1920

ttcttaggaa cttgagctgt tttccatctt ttgagaccca tgctttgtct ctctcatttt 1980ttcttaggaa cttgagctgt tttccatctt ttgagaccca tgctttgtct ctctcatttt 1980

taattctggg tctctttctt ctcttgtcct gattttttaa atgtgcttct tttttgcttc 2040taattctggg tctctttctt ctcttgtcct gattttttaa atgtgcttct tttttgcttc 2040

ttacaaccac cctctaaacc attcatcatg cttggtttgc ttttgcttct cctttcacag 2100ttacaaccac cctctaaacc attcatcatg cttggtttgc ttttgcttct cctttcacag 2100

gtttcaatca cgcaaaacaa tgctgcaatg atgctgtact tggagtttct tctgtgaccc 2160gtttcaatca cgcaaaacaa tgctgcaatg atgctgtact tggagtttct tctgtgaccc 2160

cttttttcct tccttcaaca atcaacccac cagagaaaag tgtctcagat tttgagacta 2220cttttttcct tccttcaaca atcaacccac cagagaaaag tgtctcagat tttgagacta 2220

ctttcaactt tcaaaacaaa gtggatggga tcttcatctt atataaccac acatcaatca 2280ctttcaactt tcaaaacaaa gtggatggga tcttcatctt atataaccac acatcaatca 2280

tttgtgctac ttctccaatt ttctttagag atgaaatgaa gagctaagca gacaagacaa 2340tttgtgctac ttctccaatt ttctttagag atgaaatgaa gagctaagca gacaagacaa 2340

gtttatttgt ttgttgctg atg ggt ctt ggg ttc aag atg cag cag age cac 2392gttatttgt ttgttgctg atg ggt ctt ggg ttc aag cg cag age cac 2392

Met Gly Leu Gly Phe Lys Met Gln Gln Ser His15 10Met Gly Read Gly Phe Lys Met Gln Gln Ser His15 10

cac cct gtg gct ttg aag tta cat gag caa gct ggg age cag aga aag 2440His Pro Val Ala Leu Lys Leu His Glu Gln Ala Gly ser Gln Arg Lys15 20 25cac cct gtg gt ttg aag tta cat gag caa gct ggg age cag aga aag 2440His Pro Val Wing Leu Lys Leu His Glu Gln Wing Gly Be Gln Arg Lys15 20 25

ttc act ttc att cag aac ttc aga aac tgg ttt cta ccc ctt ctg ttt 2488Phe Thr Phe Ile Gln Asn Phe Arg Asn Trp Phe Leu Pro Leu Leu Phe35 40ttc act ttc att cag aac ttc aga aac tgg ttt cta ccc ctt ctg ttt 2488Phe Thr Phe Ile Gln Asn Phe Arg Asn Trp Phe Leu Pro Leu Phe35 40

gta tgg ttc att gtt atg gct gea ttt ggt gcc tgc ate tac cat aaa 2536Val Trp Phe Ile Val Met Ala Ala Phe Gly Ala Cys Ile Tyr His Lys45 50 55atg gat gct gaa act aaa gtc aga agg aaa gag gtg ctg ggt age ctc 2584Met Asp Ala Glu Thr Lys Val Arg Arg Lys Glu Val Leu Gly Ser Leu60 65 70 75gta tgg ttc att gtt atg gct gea ttt ggt gcc tgc till tac cat aaa 2536Val Trp Phe Ile Val Met Wing Ala Phe Gly Ala Cyr Ile Tyr His Lys45 50 55atg gat gct gaa act aaa gag gtg ctg 25g Asp Wing Glu Thr Lys Val Arg Arg Lys Glu Val Leu Gly Ser Leu60 65 70 75

tgt gat caa agg gct aga atg cta caa gac caa ttc agt gtc agt gtc 2632Cys Asp Gln Arg Ala Arg Met Leu Gln Asp Gln Phe Ser Val Ser Valtgt gat caa agg gct aga atg cta caa gac caa ttc agt gtc agt gtc 2632Cys Asp Gln Arg Ala Arg Met Leu Gln Asp Gln Phe Ser Val Ser Val

80 85 9080 85 90

aac cat gtc cat gcc ctt gcc ate ctt gtt tca acc ttc cat tac tac 2680Asn His Val His Ala Leu Ala Ile Leu Val Ser Thr Phe His Tyr Tyr95 100 105aac cat gtc cat gcc ctt gcc until ctt gtt tca acc ttc cat tac tac 2680Asn His Val His Ala Leu Ala Ile Leu Val Ser Thr Phe His Tyr Tyr95 100 105

aga aat act tca gcc att gac cag gtttgtgctt gattttcctt tccttgaagc 2734Arg Asn Thr Ser Ala Ile Asp Gln110 115aga aat act tca gcc att gac cag gtttgtgctt gattttcctt tccttgaagc 2734Arg Asn Thr Ser Ala Ile Asp Gln110 115

attttttagt tggaggctca atttcttttt ctgatttgat tctggcctta aaaattagaa 2794tcaattgtag aaggatttcc aaacatgccc attttggaaa ttggtgcatc tgatagtatc 2854atgtttagat cagtttcttt ttcctcagaa ttgattttgg gcttaaaatc aattgtggaa 2914ggatattcat tagtaatttg gatattgttg catcatatgg ttctatctag ttacatcatt 2974tttttccact ctgattgcat gtatctttct cctgttcttt tccctatcag gaa acc 3030attttttagt tggaggctca atttcttttt ctgatttgat tctggcctta aaaattagaa 2794tcaattgtag aaggatttcc aaacatgccc attttggaaa ttggtgcatc tgatagtatc 2854atgtttagat cagtttcttt ttcctcagaa ttgattttgg gcttaaaatc aattgtggaa 2914ggatattcat tagtaatttg gatattgttg catcatatgg ttctatctag ttacatcatt 2974tttttccact ctgattgcat gtatctttct cctgttcttt tccctatcag gaa acc 3030

Glu ThrGlu thr

ttt gea gaa tac acg gcc agg aca gea ttt gaa cgg cca tta atg agt 3078Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe Glu Arg Pro Leu Met Serttt gea gaa tac acg gcc agg aca gea ttt gaa cgg cca tta atg agt 3078Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe Glu Arg Pro Leu Met Ser

120 125 130120 125 130

ggg gtg gcc tat gea cag aga gtg gtt cac tca gag aga gaa aga ttt 3126Gly Val Ala Tyr Ala Gln Arg Val Val His Ser Glu Arg Glu Arg Phe 135 140 145ggg gtg gcc tat gea cag aga gtg gtt cac tca gag aga gaa aga ttt 3126Gly Val Wing Tyr Wing Gln Arg Val Val His Ser Glu Arg Glu Arg Phe 135 140 145

gag aag caa cat ggg tgg gtt ata aag aca atg gaa aga gtg cct tca 3174Glu Lys Gln His Gly Trp Val Ile Lys Thr Met Glu Arg Val Pro Ser150 155 160 165gag aag caa cat ggg tgg gtt ata aag aca atg gaa aga gtg cct tca 3174Glu

ggg gtt agg gat gag tat gea gea gtg ata ttt gea cag gaa act gtc 3222Gly Val Arg Asp Glu Tyr Ala Ala Val Ile Phe Ala Gln Glu Thr Valggg gtt agg gat gag tat gea gea gtg ata ttt gea cag gaa act gtc 3222Gly Val Arg Asp Glu Tyr Ala Val Wing Ile Phe Ala Gln Glu Thr Val

170 175 180170 175 180

tet tac ctt gaa tet att gat atg atg tet ggg gag gtaaatgtca 3268Ser Tyr Leu Glu ser lie Asp Met Met Ser Gly Glutet tac ctt gaa tet att gat atg tet ggg gag gtaaatgtca 3268Ser Tyr Leu Glu ser lie Asp Met Met Ser Gly Glu

185 190185 190

acacttgtga attaattgta aaactcagaa gctactcaga gaagctcttc cccagaattg 3328gttctgcctt tagaataaat tgtacatgga tttgaccaca ttttctcatt tgcatgatgc 3388ag gag gac cga gag aac att ttg agg gct aga gcc act ggg aaa gct 3435Glu Asp Arg Glu Asn Ile Leu Arg Ala Arg Ala Thr Gly Lys Ala195 200 205gag gac cg gg gg lg gg gg lg gg gg gg gg lg gg gg gg lg gg gg lg lg gg lg gg gg gg

gtt ctg act age cct ttc aga ctg ctg gat tet cat cat ctt ggc gtg 3483Val Leu Thr Ser Pro Phe Arg Leu Leu Asp Ser His His Leu Gly vai210 215 220gtt ctg act age cct ttc aga ctg cat ctg gat tet cat cat ctt ggc gtg 3483Val Leu Thr Be Pro Phe Arg Leu Asp Ser His His Leu Gly vai210 215 220

gtt cta aca ttt cct gtt tat aaa tet aag ctc cct cca gag cca acg 3531Val Leu Thr Phe Pro vai Tyr Lys Ser Lys Leu Pro Pro Glu Pro Thr225 230 235 240gtt cta aca ttt cct gtt tat aaa tet aag ctc cct cca gag cca acg 3531Val Read Thr Phe Pro Go Tyr Lys Ser Lys Pro Le Glu Pro Thr225 230 235 240

acg gaa gag gtc att aaa gcc ata gea gg gtatgtcctc atttcacttt 3580acg gaa gag gtc att aaa gcc ata gea gg gtatgtcctc atttcacttt 3580

Thr Glu Glu Val Ile Lys Ala Ile Ala GlyThr Glu Glu Val Ile Lys Wing Ile Wing Gly

245245

tcttgccaaa accagacttc tatttggttg tgtttccgta ggctatgact gatatgtagt 3640ttcaactcag ttagactata atataaaccc ttcatgtggc ccttacccaa cagcttaagc 3700ttttgggata attggttgct tgacaaactc cttccgtaga aaacttggtt agctttggtt 3760ctatgtgggc tttatgtttt ccctgagctt atgtaatagc atgatgtgtt taatgtactt 3820tttaatggaa acag a tat att gga gga tcc ttt gat gtt gag tcc ctt gtg 3871Tyr Ile Gly Gly ser Phe Asp vai Glu Ser Leu Val255 260tcttgccaaa accagacttc tatttggttg tgtttccgta ggctatgact gatatgtagt 3640ttcaactcag ttagactata atataaaccc ttcatgtggc ccttacccaa cagcttaagc 3700ttttgggata attggttgct tgacaaactc cttccgtaga aaacttggtt agctttggtt 3760ctatgtgggc tttatgtttt ccctgagctt atgtaatagc atgatgtgtt taatgtactt 3820tttaatggaa acag tat att gga gga tcc ttt gat GTT gag tcc ctt gtg 3871Tyr Ile Gly Gly be Phe Asp will Glu Ser Leu Val255 260

gag aat tta ttt ggt caa ctt gct ggt aac caa gea att ttg gtg aag 3919Glu Asn Leu Phe Gly Gln Leu Ala Gly Asn Gln Ala Ile Leu Val Lys265 270 275gag aat tta ttt ggt caa ctt gct ggt aac caa gea att ttg gtg aag 3919Glu Asn Leu Phe Gly Gln Leu Ala Gly Asn Gln Ala Ile Leu Val Lys265 270 275

gta tat gat ata aca aac tet age gac ccc cta ate atg tat ggc age 3967Val Tyr Asp Ile Thr Asn Ser Ser Asp Pro Leu Ile Met Tyr Gly Ser280 285 290gta tat gat ata aca aac tet age gac ccc cta until atg tat ggc age 3967Val Tyr Asp Ile Thr Asn Ser Ser Asp Pro Leu Ile Met Tyr Gly Ser280 285 290

caa tat gaa gag ggt gat atg tet ctt gtc cat gaa agt aag ctt gat 4015Gln Tyr Glu Glu Gly Asp Met Ser Leu Val His Glu Ser Lys Leu Asp295 300 305 310ttt gga gat cca tac agg aaa cat cac atg ate tgt ag gtgggtgctt 4063Phe Gly Asp Pro Tyr Arg Lys His His Met Ile Cys Argcaa tat gaa gag ggt gat atg tet ctt gtc cat gaa agt aag ctt gat 4015Gln Tyr Glu Glu Gly Asp Met Ser Leu Val His Glu Ser Lys Leu Asp295 300 305 310ttt gga gat cca tac agg aaa cat cac atg till tgt ag gtgggtgcly 40 Asp Pro Tyr Arg Lys His Met Ile Cys Arg

315 320315 320

ctagttattg ttgtagtaca ttttctgagt ttcagtggtt tatcaattat cagcagattc 4123ttatgatcaa tttttttaac ag a tat cac caa cag gea cca aca aat tgg 4173ctagttattg ttgtagtaca ttttctgagt ttcagtggtt tatcaattat cagcagattc 4123ttatgatcaa tttttttaac ag to tat ca ca ca cag gea cca aca aat tgg 4173

Tyr His Gln Gln Ala Pro Thr Asn Trp325 330Tyr His Gln Gln Pro Wing Thr Asn Trp325 330

ata gea tat acc acg gea ttc cta ttc ttt gtg att ctt tgt tta gtg 4221Ile Ala Tyr Thr Thr Ala Phe Leu Phe Phe vai Ile Leu Cys Leu vai 335 340 345ata gea tat acc acg gea ttc cta ttc ttt gtg att ctt tgt tta gtg 4221Ile Wing Tyr Thr Thr Wing Phe Leu Phe Phe Go Ile Leu Cys Leu Go 335 340 345

ggt tac att tta tat gct gct gga act cac att gtc aag gta gaa gat 4269Gly Tyr Ile Leu Tyr Ala Ala Gly Thr His Ile Val Lys Val Glu Asp350 355 360ggt tac att tta tat gct gct gga act cac att gtc aag gta gaa gat 4269Gly Tyr Ile Leu Tyr Wing Gly Thr His Ile Val Lys Val Glu Asp350 355 360

gat tac aat gea atg cag gat tta aaa gtc aaa gea gaa gea gct gat 4317gat tac aat gea atg cag gat tta aaa gtc aaa gea gaa gea gct gat 4317

Asp Tyr Asn Ala Met Gln Asp Leu Lys vai Lys Ala Glu Ala Ala AspAsp Tyr Asn Ala Met Gln Asp Leu Lys Goes Lys Ala Glu Ala Asp Wing

365 370 375 380365 370 375 380

att gcc aag tca cag gtacttttca tgacatgtta gcactgttcg ttatttcctt 4372Ile Ala Lys Ser Gln385att gcc aag tca cag gtacttttca tgacatgtta gcactgttcg ttatttcctt 4372Ile Ala Lys Ser Gln385

gaattgcata ctgatcacta gaaactgaaa atttgttatt aatgtcag ttt cta gct 4429gaattgcata ctgatcacta gaaactgaaa atttgttatt aatgtcag ttt cta gct 4429

Phe Leu AlaPhe Leu Wing

acc gtc tet cat gaa att aga act ccc atg aat gga att tta g 4472acc gtc tet cat gaa att aga act ccc atg aat gga att tta g 4472

Thr vai Ser His Glu Ile Arg Thr Pro Met Asn Gly Ile Leu 390 395 400Thr Will Be His Glu Ile Arg Thr Pro Met Asn Gly Ile Leu 390 395 400

gtaactttaa gattetetet cgttctcttt ccactgaaaa gcaacacatg ctttcattcc 4532atacctgata ctttcccatt agtgatgcta tcgttaaact ccttgtcact gtag ga 4588gtaactttaa gattetetet cgttctcttt ccactgaaaa gcaacacatg ctttcattcc 4532atacctgata ctttcccatt agtgatgcta tcgttaaact ccttgtcact gtag ga 4588

GlyGly

atg ctt ggt ctg ctt tta cgc aca gaa ttg agt tca aca caa aga gac 4636Met Leu Gly Leu Leu Leu Arg Thr Glu Leu Ser Ser Thr Gln Arg Asp405 410 415tat gct cag act gct caa gca tgt ggg aag gca cta ata gca tta ata 4684atg ctt ggt ctg ctt tta cgc aca gaa ttg agt tca aca caa aga 4636Met Leu Gly Leu Leu Leu Arg Thr Glu Leu Being Thr Gln Arg Asp405 410 415tat gct cag act gct ggg aag gta cta 468

Tyr Ala Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu lie Ala Leu Ile420 425 430 435Tyr Wing Gln Thr Wing Gln Cys Wing Gly Lys Wing Leu lie Wing Leu Ile420 425 430 435

aat gag gtg ctt gac cga gct aaa att gaa gca ggc aaa tta gag cta 4732Asn Glu Val Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys Leu Glu Leuaat gag gtg ctt gac cga gct aaa att gaa gca ggc aaa tta gag cta 4732Asn Glu Val Leu Asp Arg Wing Lys Ile Glu Wing Gly Lys Leu Glu Leu

440 445 450440 445 450

gaa gca gtt cca ttt gac ctt cgt tcc ata ctt gac gat gtc ctt tct 4780Glu Ala vai Pro Phe Asp Leu Arg ser Ile Leu Asp Asp vai Leu Ser455 460 465gaa gca gtt cca ttt gac ctt cgt tcc ata ctt gac gat gtc ctt tct 4780Glu Wing Go Pro Phe Asp Leu Arg Be Ile Leu Asp Asp Go Leu Ser455 460 465

ctt ttt tct gag aag tca aga cac aaa ggc tta gag gtacgtttag 4826ctt ttt tct gag aag tca aga cac aaa ggc tta gag gtacgtttag 4826

Leu Phe Ser Glu Lys Ser Arg His Lys Gly Leu GluLeu Phe Be Glu Lys Be Arg His Lys Gly Leu Glu

470 475470 475

tcattgctaa atctgttgtg aagtcctgta caagtggcgt aatttcatag tcctatttcc 4886ttttcttcaa ttgatgatct tatttcatat tcctctcgtg tttctctctt tctatgttgc 4946catgtcgttg ggcggtgatg ttcctactct gatcccaaat tcctgatgtg caattattcc 5006gacttggact tcaatggttt gggaagtata g ctg gca gtg ttt gtt tct gac 5058tcattgctaa atctgttgtg aagtcctgta caagtggcgt aatttcatag tcctatttcc 4886ttttcttcaa ttgatgatct tatttcatat tcctctcgtg tttctctctt tctatgttgc 4946catgtcgttg ggcggtgatg ttcctactct gatcccaaat tcctgatgtg caattattcc 5006gacttggact tcaatggttt gggaagtata g ctg gca gtg gtt tct gac ttt 5058

Leu Ala vai Phe Val Ser Asp480 485Leu Wing goes Phe Val Ser Asp480 485

aaa gtt ccg gat ata gtt atg ggc gat cct ggg cga ttc aga caa ata 5106Lys vai Pro Asp Ile Val Met Gly Asp Pro Gly Arg Phe Arg Gln Ileaaa gtt ccg gat ata gtt atg ggc gat cct ggg cga ttc aga caa ata 5106Lys goes Pro Asp Ile Val Met Gly Asp Pro Gly Arg Phe Arg Gln Ile

490 495 500490 495 500

gtg aca aat ctt gtt gga aac tct gtt aag gttagtggaa ttttcaaact 5156gtg aca aat ctt gtt gga aac tct gtt aag gttagtggaa ttttcaaact 5156

vai Thr Asn Leu Val Gly Asn ser Val Lys505 510Will Thr Asn Read Val Gly Asn Be Val Lys505 510

ttatttgcct aatgttgtgt gcaagttgtg tgttggaaat gcgtcctttt aacgttataa 5216aatcgtacaa gttcgtattc tccattgtat acaataactt attagcaaag tacttgttga 5276tatcattact gattaacttt aatatcttgc ag ttc act gag cga ggt cat ata 5329ttatttgcct aatgttgtgt gcaagttgtg tgttggaaat gcgtcctttt aacgttataa 5216aatcgtacaa gttcgtattc tccattgtat acaataactt attagcaaag tacttgttga 5276tatcattact gattagg cggtgatta

Phe Thr Glu Arg Gly His Ile515Phe Thr Glu Arg Gly His Ile515

ttt gtt aaa gtc cat tta gct gaa aaa aga cag tgc aca atg aat gga 5377Phe vai Lys vai His Leu Ala Glu Lys Arg Gln Cys Thr Met Asn Gly520 525 530 535aaa tgt gag act ttt cta aat gga ggc tgt gat gat gtt ttg cat gta 5425Lys Cys Glu Thr Phe Leu Asn Gly Gly Cys Asp Asp Val Leu His Valttt gtt aaa gtc cat tta gct gaa aaa aga cag tgc aca atg aat gga 5377Phe will Lys will His Leu Ala Glu Lys Arg Gln Cys Thr Met Asn Gly520 525 530 535aaa tgt gag act cat tt gt gat gat catg tgt gat gat 5425Lys Cys Glu Thr Phe Leu Asn Gly Gly Cly Asp Asp Val Leu His Val

540 545 550540 545 550

tct ggc agt tat aat ttg aaa acc ctt agt gga tat gaa gcc gct gat 5473Ser Gly Ser Tyr Asn Leu Lys Thr Leu Ser Gly Tyr Glu Ala Ala Asp555 560 565tct ggc agt tat aat ttg aaa acc ctt agt gga tat gaa gcc gct gat 5473Ser Gly Ser Tyr Asn Leu Lys Thr Leu Ser Gly Tyr Glu Wing Asp555 560 565

gaa cgg aac age tgg gat aat ttt aag cat cat att gct gac gaa gaa 5521Glu Arg Asn Ser Trp Asp Asn Phe Lys His His Ile Ala Asp Glu Glu570 575 580cat cgg aac age tgg gat aat ttt aag cat cat att gct gac gaa gaa 5521Glu Arg Asn Ser Trp Asp Asn Phe Lys His Ile Ala Asp Glu Glu570 575 580

ttt ttc ttt gat gct tcg gtt aaa aag ttg gcc tct agt gaa tct tat 5569Phe Phe Phe Asp Ala Ser vai Lys Lys Leu Ala Ser Ser Glu Ser Tyrttt ttc ttt gat gct tcg gtt aaa aag ttg gcc tct agt gaa tct tat 5569Phe Phe Phe Asp Ala Ser Go Lys Lys Leu Ala Ser Be Glu Ser Tyr

585 590 595585 590 595

gag caa gtc acc ttg atg gtc age gtg gag gac act gga att ggg att 5617Glu Gln Val Thr Leu Met vai Ser vai Glu Asp Thr Gly Ile Gly Ile 600 605 610 615gag caa gtc acc ttg atg gtc age gtg gag gac act gga att ggg att 5617Glu Gln Val Thr Leu Met Will Be Going Glu Asp Thr Gly Ile Gly Ile 600 605 610 615

tct ttc tct gcc caa gat agt att ttc atg cct ttt gtg cag gct gac 5665Ser Phe Ser Ala Gln Asp Ser Ile Phe Met Pro Phe vai Gln Ala Asptct ttc tct gcc caa gat agt att ttc atg cct ttt gtg cag gct gac 5665Ser Phe Ser Ala Gln Asp Ser Ile Phe Met Pro Phe Go Gln Ala Asp

620 625 630620 625 630

age tca acc tct cga aac tat ggg ggt acc ggg ate ggc ttg agt ate 5713 Ser Ser Thr Ser Arg Asn Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile635 640 645age tca acc tct cga aac tat ggg ggt acc ggg until ggc ttg agt until 5713 Ser Ser Thr Ser Arg Asn Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile635 640 645

agt aag tgc ttg gtt gaa ctg atg ggc ggt cag ata aac ttc ata age 5761Ser Lys Cys Leu Val Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Ser650 655 660agt aag tgc ttg gtt gaa ctg atg ggc ggt cag ata aac ttc ata age 5761Ser Lys Cys Leu Val Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Ser650 655 660

cga ccc cag gtc ggg age acg ttt tca ttc act gea gat ttc gga aca 5809Arg Pro Gln vai Gly Ser Thr Phe Ser Phe Thr Ala Asp Phe Gly Thrcga ccc cag gtg ggg age acg ttt tca ttc act gea gat ttc gga ac 5809Arg Pro Gln will Gly Be Thr Phe Be Phe Thr Wing Asp Phe Gly Thr

665 670 675665 670 675

ttt aag aaa aac tca aca act gac atg aag aaa ctt aac ttt gaa gat 5857Phe Lys Lys Asn Ser Thr Thr Asp Met Lys Lys Leu Asn Phe Glu Asp680 685 690 695att aaa aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 5857Phe

cta cct tct agt ttt aga ggt ctt aaa gcc att gtg gtt gat gga aaa 5905Leu Pro Ser Ser Phe Arg Gly Leu Lys Ala Ile Val Val Asp Gly Lys700 705 710cta cct tct agt ttt aga ggt ctt aaa gcc att gtg gtt gat gga aaa 5905Leu Pro Be Ser Phe Arg Gly Leu Lys Ala Ile Val Val Asp Gly Lys700 705 710

cct gtt aga gct gca gtg act aga tac cat ttg aag aga cta ggg ata 5953Pro Val Arg Ala Ala Val Thr Arg Tyr His Leu Lys Arg Leu Gly Ile715 720 725cct gtt aga gct gca gtg act aga tac cat ttg aag aga cta ggg ata 5953Pro Val Arg Wing Wing Val Thr Arg Tyr His Leu Lys Arg Leu Gly Ile715 720 725

caa gct aaa gtt gca att age ate aat aag gct gtt tet tta tgt ggg 6001Gln Ala Lys Val Ala Ile Ser Ile Asn Lys Ala Val Ser Leu Cys Gly730 735 740caa gct aaa gtt gca att age till aat aag gct gtt tet tta tgt ggg 6001Gln Alys Lys Val Ala Ile Ser Ile Asn Lys Ala Val Ser Leu Cys Gly730 735 740

aaa aat ggt tet ttg acc tcg gc gtaagtcttt aattaacctt tttggtttca 6054Lys Asn Gly Ser Leu Thr Ser Ala745 750aaa aat ggt tet ttg acc tcg gc gtaagtcttt aattaacctt tttggtttca 6054Lys Asn Gly Ser Leu Thr Ser Ala745 750

attatgtaga aatgtattga atgttatgat aatcagtagc attatcaact tttagtaatt 6114gttcttaaca tatgctaata gtcatatctt tctataatac tacaatactg tagccatata 6174atatctttcc tgtattggag tgagttttca aatgtttttc tgtgatattt tggaagttat 6234cttcagtttg agaactcatt tgtcattttt gcattttgtt attggatatt tggatggatc 6294tttacaaagg atgtgtggat tttgacttgt tatacacatt tcttctccat tttatattgt 6354ttgtgttatt ctttttactc ataaagaaat ttagaaactg cattgactgg ttctttttaa 6414ttacttacag atattgacat tgatattttt tgtaaatgct gtcttgacat ggtttaatta 6474cttacagact aggtttttct ttccttttct aacatgcata tccatttact tttttgacca 6534accaacatcc tcatgagtca tgacatgttg atgatttata tggttgactt gagactattt 6594agacattaaa taaccgcaaa ttccatgttg tttgtgtgtt tggttccctg ttgggtaatc 6654tcagaatcaa ttatgataga gtaaaatcaa ttttggatga gatgtgtggg tgtcattttg 6714taaacctaaa cccaaaatca attctgctat aagetagaga gagtagttga acataatcaa 6774ttgtgagatt ttgcaagtgg attgcacaac attgccctat gaaaatcact ttttgttcac 6834aaaatttatc taaacataca taacttcatt ttcaaccttt actataatcà attttacaat 6894aattaatttt acccaaaatc aattgtgaca atgagtttcc aaacacacac ttaaagacta 6954ccatttgcag aaaatatgtg atagaagact tatgtttatg tagtgtgttt cagttcattc 7014actgatttaa actactcgga ttttgcag a tta ttt cag cct gat att att ttt 7067attatgtaga aatgtattga atgttatgat aatcagtagc attatcaact tttagtaatt 6114gttcttaaca tatgctaata gtcatatctt tctataatac tacaatactg tagccatata 6174atatctttcc tgtattggag tgagttttca aatgtttttc tgtgatattt tggaagttat 6234cttcagtttg agaactcatt tgtcattttt gcattttgtt attggatatt tggatggatc 6294tttacaaagg atgtgtggat tttgacttgt tatacacatt tcttctccat tttatattgt 6354ttgtgttatt ctttttactc ataaagaaat ttagaaactg cattgactgg ttctttttaa 6414ttacttacag atattgacat tgatattttt tgtaaatgct gtcttgacat ggtttaatta 6474cttacagact aggtttttct ttccttttct aacatgcata tccatttact tttttgacca 6534accaacatcc tcatgagtca tgacatgttg atgatttata tggttgactt gagactattt 6594agacattaaa taaccgcaaa ttccatgttg tttgtgtgtt tggttccctg ttgggtaatc 6654tcagaatcaa ttatgataga gtaaaatcaa ttttggatga gatgtgtggg tgtcattttg 6714taaacctaaa cccaaaatca attctgctat aagetagaga gagtagttga acataatcaa 6774ttgtgagatt ttgcaagtgg attgcacaac attgccctat gaaaatcact ttttgttcac 6834aaaatttatc taaacataca taacttcatt ttcaaccttt actataatcà attttacaat 6894aattaatttt acccaaaa tc aattgtgaca atgagtttcc aaacacacac ttaaagacta 6954ccatttgcag aaaatatgtg atagaagact tatgtttatg tagtgtgttt cagttcattc 7014actgatttaa actactcgga ttttgcag a tta ttt cag cct gat att att ttt 70

Leu Phe Gln Pro Asp Ile Ile Phe755Read Phe Gln Pro Asp Ile Ile Phe755

gtt gag aag gac tet tgg gtt tet gga gag gat ggt ggt ate ttc aat 7115vai Glu Lys Asp Ser Trp Val Ser Gly Glu Asp Gly Gly Ile Phe Asn760 765 770 775gcg ttt aag atg cct caa atg ate ctt ctt gea acc aat ate tgt aac 7163Ala Phe Lys Met Pro Gln Met Ile Leu Leu Ala Thr Asn Ile Cys Asngtt gag aag gac tet tgg gtt tet gga gag gat ggt ggt till ttc aat 7115vai Glu Lys Asp Ser Trp Val Ser Gly Glu Asp Gly Gly Ile Phe Asn760 765 770 775gcg ttt aag atg cct caa tg acc a cg t 7163Ala Phe Lys Met Pro Gln Met Ile Read Leu Wing As Thr In Cys Asn

780 785 790780 785 790

gct gaa ttt gat aaa gcc aaa gct gea ggt ttc agt gat aca gtg ate 7211Ala Glu Phe Asp Lys Ala Lys Ala Ala Gly Phe ser Asp Thr vai Ile795 800 805gt gaa ttt gat aaa gcc aaa gct gea ggt ttc agt gat aca gtg till 7211Ala Glu Phe Asp Lys Ala Lys Ala Gly Phe Be Asp Thr Go Ile795 800 805

atg aag cca ctg aga gct agt atg ctg gct gct tgt ctt cag caa gtt 7259Met Lys Pro Leu Arg Ala Ser Met Leu Ala Ala Cys Leu Gln Gln Val810 815 820atg aag cca ctg aga gct agt atg ctg gct gct tgt ctt cag caa gtt 7259Met Lys Pro Leu Arg Ala Ser Met Leu Ala Cys Leu Gln Gln Val810 815 820

ttc ggg act ggc aag acg agg cag ttt ggg aaa gac atg tcg aat ggt 7307Phe Gly Thr Gly Lys Thr Arg Gln Phe Gly Lys Asp Met Ser Asn Glyttc ggg act ggc aag acg agg cag ttt ggg aaa gac atg tcg aat ggt 7307Phe Gly Thr Gly Lys Thr Arg Gln Phe Gly Lys Asp Met Ser Asn Gly

825 830 835825 830 835

tet tca gta cga age ctt ctt tgc gga aag aaa ate tta gtg gtt gat 7355Ser Ser Val Arg Ser Leu Leu Cys Gly Lys Lys Ile Leu vai vai Asp840 845 850 855tet tca gta cga age ctt ctt tgc gga aag aaa till tta gtg gtt gat 7355Ser Ser Val Arg Ser Leu Cys Gly Lys Lys Ile Leu Go Go Asp840 845 850 855

gat aat ttg gtg aac cga agg gtc gcc gcc ggc gcg ttg aaa aac ttt 7403Asp Asn Leu Val Asn Arg Arg Val Ala Ala Gly Ala Leu Lys Asn Phegat aat ttg gtg aac cga agg gtc gcc gcc gcc gcg ttg aaa aac ttt 7403Asp Asn Leu Val Asn Arg Arg Val Wing Ala Gly Wing Leu Lys Asn Phe

860 865 870860 865 870

gga gct gat gtc aaa tgt gea gea agt ggc aaa gct gct ctt gaa atg 7451Gly Ala Asp Val Lys Cys Ala Ala Ser Gly Lys Ala Ala Leu Glu Met875 880 885gga gct gat gtc aaa tgt gea gea agt ggc aaa gct gct ctt gaa atg 7451Gly Wing Asp Val Lys Wing Wing Wing Ser Gly Lys Wing Wing Leu Glu Met875 880 885

ctt caa tat cct cac gat ttc gat gct tgc ttc atg gat att caa atg 7499Leu Gln Tyr Pro His Asp Phe Asp Ala Cys Phe Met Asp Ile Gln Met890 895 900ctt caa tat cct cac gat ttc gat gct tgc ttc atg gat att caa atg 7499Leu Gln Tyr Pro His Asp Phe Asp Ala Cys Phe Met Asp Ile Gln Met890 895 900

cca gaa atg gat gg gtatgcttac tggcactgac taatacatgt tttttgccaa 7553Pro Glu Met Asp Gly905cca gaa atg gat gg gtatgcttac tggcactgac taatacatgt tttttgccaa 7553Pro Glu Met Asp Gly905

cttaatatat tactctttca atattcgttg tgttattaga agatcatata gattaattta 7613taaattttct tttagcaaaa ccttatcaat taagtgtgta gaaaagtcag tctcacatta 7673tggtcaaata agtgttaggg caagcttcac ctcaaagcta gctatttggg tagatttagg 7733cctaacccga attctaagat ggtatcagag tctatcctag atctttttat tggaaaccac 7793ccgtatatga gcaactcgta gatatteatt cttgaaagtt gcacgctcca tatgtccatt 7853cctaggtgcg agagagaagt ctcactttga ctagagatat gattaaaaaa atatttataa 7913agggttgagc aatcctcacc teagagetaa gcttttgggg taaagttagg cctaactcga 7973actctaataa agtgtttagc tggtgtgtca actgtcaata tgaaatcttt tgcaatttac 8033tatgcattca cttacctact ttattgaagc ttattgacaa tttgtgcaga agcatcatta 8093attaggaaca tgttagctat acaagttatg atgtttttgt atageatate atgttccaac 8153cttccaataa caaaatatgt ggttcaagtg tgagaatata taggttaaac aataaagtat 8213tgagttaaca gaaatctaaa cacacgctgt cactagctct tcatattgag acatgcatgg 8273gatttgacaa aacatctgaa taaatatttg cag g ttt gag gcc act cgt cga 8325cttaatatat tactctttca atattcgttg tgttattaga agatcatata gattaattta 7613taaattttct tttagcaaaa ccttatcaat taagtgtgta gaaaagtcag tctcacatta 7673tggtcaaata agtgttaggg caagcttcac ctcaaagcta gctatttggg tagatttagg 7733cctaacccga attctaagat ggtatcagag tctatcctag atctttttat tggaaaccac 7793ccgtatatga gcaactcgta gatatteatt cttgaaagtt gcacgctcca tatgtccatt 7853cctaggtgcg agagagaagt ctcactttga ctagagatat gattaaaaaa atatttataa 7913agggttgagc aatcctcacc teagagetaa gcttttgggg taaagttagg cctaactcga 7973actctaataa agtgtttagc tggtgtgtca actgtcaata tgaaatcttt tgcaatttac 8033tatgcattca cttacctact ttattgaagc ttattgacaa tttgtgcaga agcatcatta 8093attaggaaca tgttagctat acaagttatg atgtttttgt atageatate atgttccaac 8153cttccaataa caaaatatgt ggttcaagtg tgagaatata taggttaaac aataaagtat 8213tgagttaaca gaaatctaaa cacacgctgt cactagctct tcatattgag acatgcatgg 8273gatttgacaa aacatctgaa taaatatttg g cag gag ttt gcc cgt cga act 8325

Phe Glu Ala Thr Arg Arg910Phe Glu Wing Thr Arg Arg910

att cgg atg atg gaa aga gag gca agt gag cag ctg aaa agt gaa tct 8373Ile Arg Met Met Glu Arg Glu Ala Ser Glu Gln Leu Lys Ser Glu Ser915 920 925 930att cgg atg atg gaa aga gag gca agt gag cag ctg aaa agt gaa tct 8373Ile Arg Met Met Glu Arg Glu Ala Ser Glu Gln Leu Lys Ser Glu Ser915 920 925 930

ggt gaa gaa aat ggt aag aaa agt gag ttc cac atg cct ata ttg gcc 8421Gly Glu Glu Asn Gly Lys Lys Ser Glu Phe His Met Pro Ile Leu Alaggt gaa gaa aat ggt aag aaa agt gag ttc cac atg cct ata ttg gcc 8421Gly Glu Glu Asn Gly Lys Lys Ser Glu Phe His Met Pro Ile Leu Ala

935 940 945935 940 945

atg aca gct gat gta ate cat gct aca tat gat aag tgc tta aat tgt 8469Met Thr Ala Asp vai Ile His Ala Thr Tyr Asp Lys Cys Leu Asn Cys950 955 960atg aca gct gat gta until cat gct aca tat gat aag tgc tta aat tgt 8469Met Thr Ala Asp Goes Ile His Ala Thr Tyr Asp Lys Cys Leu Asn Cys950 955 960

ggg atg gat gga tac gtc tca aag cct ttt gaa gaa gag aat ctc tat 8517Gly Met Asp Gly Tyr Val Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyrggg atg gat gga tac gtc tca aag cct tt gaa gaa gag aat ctc tat 8517Gly Met Asp Gly Tyr Val Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr

965 970 975965 970 975

caa gca gtt gca aag ttc ttc aag tcc aag cct gct tca gac tca tg 8564Gln Ala Val Ala Lys Phe Phe Lys Ser Lys Pro Ala Ser Asp Ser980 985 990caa gca gtt gca aag ttc ttc aag tcc aag cct gct tca gac tca tg 8564Gln Val Wing Ala Lys Phe Phe Lys Ser Lys Pro Ala Ser Asp Ser980 985 990

acactgctta ttctgcagaa caggtcaacc aacttttgat tgagaaacat ttagtgttag 8624catgtttgga tcaacttctc ccagcatcaa ttctgaaact cagaagctac tcatagaagc 8684ttctagccag aattgatttt ggcttcagaa gctactaatg gttttatgta gagagcaaaa 8744gttggtttcc aagatatgcg aggatccatg atgatacaca cgtctgaagt gatcatttct 8804aaccagttga agtttcactc gacgtgattt gaatccaagt aaatgcatac cacataatta 8864tccatcccca tcttgtgtac agattctccc aagggataag aaaatttatg taaattcaat 8924ttttttcttt tgcatctcaa tacttccctg ttagaacttt ttccctatga ttatttccac 8984ttttcatttt caattatatt ttttgtaaaatttttttaat tcagatgaga aggttggtgctgtgatataa atatttgtct tatttttagaatagcatttt tggagaatga tatcataattaaatcaatat taatgcatag caagtagctttttttttcta aactaattac aatgtattattatcatgttt ggattagttt ttcttagaatatttattctt caaagttgat tttggcaatcttgtttctta ttattgtgat tttagctcatatttgatgtg acgttgactt gtgaagcacgttgtcagcac tatgttgcta ttataagtgacataaacttt tttattaaaa gtaacagaattggaaaatta tcagaaccag tatggagctgagagtgaaat attggaaacc accaatgttgatgatggaag aaccactgct gcagatgctaaccacaagaa aggaatatta gagaagatcaaagatcacaa atagctagct agctttgtttaataataata gtatgtgtgt tgctacatgcgttaataact tgctttcaat tttgcttcctatttcactat tggtttctaa tcaatctaaattttaattaa agagtaaatg ttgaatattgtttttgcttc atcggaaaat tatttacattttcgttaaaa atttcataat taattttttaaacagaccta tctttgtatg tatggcattcaattccccac tactatacat aaaatgatacagtcatattg gtaaattgat ttcagttaaaagcttctatg tcaaaatagt taattcctatttattcaaac atctcaactc actttgactaaaataaatta tattcaaatt aaatgtgataagggaaatat ggaacaaaga gagctgcatgcaaagaagtt gtaatcacaa ttaggacaatcagggaaagt ctatagatcc actgtgctctacactgctta ttctgcagaa caggtcaacc aacttttgat tgagaaacat ttagtgttag 8624catgtttgga tcaacttctc ccagcatcaa ttctgaaact cagaagctac tcatagaagc 8684ttctagccag aattgatttt ggcttcagaa gctactaatg gttttatgta gagagcaaaa 8744gttggtttcc aagatatgcg aggatccatg atgatacaca cgtctgaagt gatcatttct 8804aaccagttga agtttcactc gacgtgattt gaatccaagt aaatgcatac cacataatta 8864tccatcccca tcttgtgtac agattctccc aagggataag aaaatttatg taaattcaat 8924ttttttcttt tgcatctcaa tacttccctg ttagaacttt ttccctatga ttatttccac 8984ttttcatttt caattatatt ttttgtaaaatttttttaat tcagatgaga aggttggtgctgtgatataa atatttgtct tatttttagaatagcatttt tggagaatga tatcataattaaatcaatat taatgcatag caagtagctttttttttcta aactaattac aatgtattattatcatgttt ggattagttt ttcttagaatatttattctt caaagttgat tttggcaatcttgtttctta ttattgtgat tttagctcatatttgatgtg acgttgactt gtgaagcacgttgtcagcac tatgttgcta ttataagtgacataaacttt tttattaaaa gtaacagaattggaaaatta tcagaaccag tatggagctgagagtgaaat attggaaacc accaatgttgatgatggaag aaccactgct gcagatgctaaccacaagaa aggaatatta gagaagat caaagatcacaa atagctagct agctttgtttaataataata gtatgtgtgt tgctacatgcgttaataact tgctttcaat tttgcttcctatttcactat tggtttctaa tcaatctaaattttaattaa agagtaaatg ttgaatattgtttttgcttc atcggaaaat tatttacattttcgttaaaa atttcataat taattttttaaacagaccta tctttgtatg tatggcattcaattccccac tactatacat aaaatgatacagtcatattg gtaaattgat ttcagttaaaagcttctatg tcaaaatagt taattcctatttattcaaac atctcaactc actttgactaaaataaatta tattcaaatt aaatgtgataagggaaatat ggaacaaaga gagctgcatgcaaagaagtt gtaatcacaa ttaggacaatcagggaaagt ctatagatcc actgtgctct

ttggtctcca tcctataagg tttgcttagt 9044ttataatgtg tatacctttt tagcagtact 9104acttaggaga taataagatc gtaggagtaa 9164ataatcatct acattctgca gttataaaaa 9224taagaagtat gatctattta agtattgaat 9284tgctgggata tgcgcttttt actcgataga 9344ccattttgga accgaaaaaa ttatcacata 9404aaggtaaaca ctcataaaag ctaaagggag 9464tgtggttttt caccgtgtat ataaacatgc 9524tggctatcca ccagcaatct caagtaacac 9584caaggcagtg accacacccc tctatatatg 9644caaaagggaa gttttctaca aattaattaa 9704ttcctgctag cagggctccg gttcagagcc 9764gcggtgctac acaagtcata agaaatgatt 9824ctgttgttgg tgagagcgat ctgcagcatc 9884aggagaagct tcctggaaca caccaccatc 9944agtttatttt gcttctatca ttataaatgt 10004atgtgtatgt atcgtttagt cagctagttt 10064tacaagccta tttatgcaat gtatttggtc 10124ttagtacatg gtcctcttcc ttttagtgcc 10184tttttagtca ttctatttac atttttattt 10244tttattaagt gtgtgtttag attttcgttt 10304taacagtgat gtatgcagct aaggcatcct 10364tgggcgggtc ggatttattt ttagccccat 10424ctgctagtaa aataatttaa agtgtttcga 10484aatcgaagtt tgttagctac taaagacggt 10544attttataat tggatcattt tttctaaaat 10604taatcacttt ataattaatt ttgttgaatt 10664atacccactg aatgtgattg gaatgccatc 10724tggaagggct tgttatgtct gcaaaacaga 10784tgccaaagat gtggggtcac attgacttgt 10844taaagctaga acaattaaca taaaatttct 10904gccaattaag gctttcactt cctatccctagacagttgtg gtgatgttat aatcaactgctgtctatata aacttttttg tcattattaaagcttttgtc ttgtgctgaa agttgactgtattgattgtg gtgtttttat gcgacaactggaggcaatta caaacaaaat acagctattaatttaaatgt gattttgaaa tcttatattgaaagtgacag aacaagtaga tggtaaacaaaagaccatca atgtcaacac ttccttggagctagacaaaa tgtagccaaa atgaagtatgctagattcca agttattgaa ggaaccagcatgaaagagga ttgaataaaa gctgaaccaacattagaatc tgttttaact tccttgtacattggtaacat gatgagtagc tcatgaatctgtagagaaga atgagaacta catgtgagcatgttgctgca atcctggatc agtacgcatagttgctctaa tatgttatga atattcaagatcacataact attaaattat tttaatcttatctcatataa aataattatt ctgatacaatggatctgagg atgtatcata tgagaaatgtcaaccgttag atttaatcaa cggaccagataaaaatgtca tgtgactttc ttaaaaagtccatgattaga tttaacggtc atgatttattttttctgata tgtatatttg taacataaaaatttcaccaa atacaatatg atgtgaaaaagcacaatcgg ccctaaaaaa tggtcaattttgagtattca tgcaatatat gatttcaaaatgcaccatgt gtaatatgta agagatcaaatgatatggga aacattttta tagataattttttcggcgaa gttggtgctt ttttttctttattggagttt aatccatttt tctatcatgattggtctcca tcctataagg tttgcttagt 9044ttataatgtg tatacctttt tagcagtact 9104acttaggaga taataagatc gtaggagtaa 9164ataatcatct acattctgca gttataaaaa 9224taagaagtat gatctattta agtattgaat 9284tgctgggata tgcgcttttt actcgataga 9344ccattttgga accgaaaaaa ttatcacata 9404aaggtaaaca ctcataaaag ctaaagggag 9464tgtggttttt caccgtgtat ataaacatgc 9524tggctatcca ccagcaatct caagtaacac 9584caaggcagtg accacacccc tctatatatg 9644caaaagggaa gttttctaca aattaattaa 9704ttcctgctag cagggctccg gttcagagcc 9764gcggtgctac acaagtcata agaaatgatt 9824ctgttgttgg tgagagcgat ctgcagcatc 9884aggagaagct tcctggaaca caccaccatc 9944agtttatttt gcttctatca ttataaatgt 10004atgtgtatgt atcgtttagt cagctagttt 10064tacaagccta tttatgcaat gtatttggtc 10124ttagtacatg gtcctcttcc ttttagtgcc 10184tttttagtca ttctatttac atttttattt 10244tttattaagt gtgtgtttag attttcgttt 10304taacagtgat gtatgcagct aaggcatcct 10364tgggcgggtc ggatttattt ttagccccat 10424ctgctagtaa aataatttaa agtgtttcga 10484aatcgaagtt tgttagctac taaagacggt 10544attttataat tggatcattt tttcta AAAT 10604taatcacttt ataattaatt ttgttgaatt 10664atacccactg aatgtgattg gaatgccatc 10724tggaagggct tgttatgtct gcaaaacaga 10784tgccaaagat gtggggtcac attgacttgt 10844taaagctaga acaattaaca taaaatttct 10904gccaattaag gctttcactt cctatccctagacagttgtg gtgatgttat aatcaactgctgtctatata aacttttttg tcattattaaagcttttgtc ttgtgctgaa agttgactgtattgattgtg gtgtttttat gcgacaactggaggcaatta caaacaaaat acagctattaatttaaatgt gattttgaaa tcttatattgaaagtgacag aacaagtaga tggtaaacaaaagaccatca atgtcaacac ttccttggagctagacaaaa tgtagccaaa atgaagtatgctagattcca agttattgaa ggaaccagcatgaaagagga ttgaataaaa gctgaaccaacattagaatc tgttttaact tccttgtacattggtaacat gatgagtagc tcatgaatctgtagagaaga atgagaacta catgtgagcatgttgctgca atcctggatc agtacgcatagttgctctaa tatgttatga atattcaagatcacataact attaaattat tttaatcttatctcatataa aataattatt ctgatacaatggatctgagg atgtatcata tgagaaatgtcaaccgttag atttaatcaa cggaccagataaaaatgtca tgtgactttc ttaaaaagtccatgattaga tttaacggtc atgatttattttttctgata tgtatatttg taacataaaaatttcaccaa atacaatatg atgtgaaaaa gcacaatcgg ccctaaaaaa tggtcaattttgagtattca tgcaatatat gatttcaaaatgcaccatgt gtaatatgta agagatcaaatgatatggga aacattttta tagataattttttcggcgaa gttggtgctt tttgtctctat a tattgctcat

<210> 5<210> 5

tctgctgtat attttaagtt tgagtaaagg 10964aggctgaggg aaaagttggg ggtcaagtgg 11024tctttaatct ttttggtttt gtaaaatgac 11084gctagctagt aaagtcatgc tggtttctta 11144tagacattgc aatgcaggca cacaagtcaa 11204ataattgacc tactaactac taaggaggga 11264gatgtaaaga ggcaatactc atcagactgt 11324gtagaagtag aagcagcaac aaattgagta 11384aagaatttca taagtgtcat gtgaaaataa 11444aaactcagtt attgaaacga gtgacactga 11504aaaggttaag tgacagtgaa gtgtgtgggt 11564tggactgaat aggcaaactt gacaatttcc 11624aaagtttctt cggtgaaatt tccagcattg 11684ggaaaccaaa cagttttgtt tgttcaagct 11744ttgttatgta ccaaaaaaac ctgtgagcat 11804gcctgagcac aaagatgttg aattgaagta 11864ttaaaacatt tttaagttat catgtgacaa 11924actattaatc attagatcta acgatcatat 11984aatatatata tatatatata tgtatgtagg 12044gatattatgt gagaacatga gaataaatca 12104taaaacaatt taatgctcac atgaccactt 12164aaatgacttc atgttttaat tttgaccgtt 12224ctcatgttct cacataaaaa catttttcat 12284cacaacataa aatactatgt aaatcagcca 12344tataatatta ctattgatat tcactcaaat 12404ttctggttaa tatatttctt ttttaaaaca 12464tatataattt ctttcaacat ttaaccaatt 12524aattatattg agttttattt atcttttaac 12584ttattcaagt tctaagttac ttcattttta 12644tcaaaccgtt tgaagatggt ttcctcatgg 12704gctc 12738<211> 3568<212> DNAtctgctgtat attttaagtt tgagtaaagg 10964aggctgaggg aaaagttggg ggtcaagtgg 11024tctttaatct ttttggtttt gtaaaatgac 11084gctagctagt aaagtcatgc tggtttctta 11144tagacattgc aatgcaggca cacaagtcaa 11204ataattgacc tactaactac taaggaggga 11264gatgtaaaga ggcaatactc atcagactgt 11324gtagaagtag aagcagcaac aaattgagta 11384aagaatttca taagtgtcat gtgaaaataa 11444aaactcagtt attgaaacga gtgacactga 11504aaaggttaag tgacagtgaa gtgtgtgggt 11564tggactgaat aggcaaactt gacaatttcc 11624aaagtttctt cggtgaaatt tccagcattg 11684ggaaaccaaa cagttttgtt tgttcaagct 11744ttgttatgta ccaaaaaaac ctgtgagcat 11804gcctgagcac aaagatgttg aattgaagta 11864ttaaaacatt tttaagttat catgtgacaa 11924actattaatc attagatcta acgatcatat 11984aatatatata tatatatata tgtatgtagg 12044gatattatgt gagaacatga gaataaatca 12104taaaacaatt taatgctcac atgaccactt 12164aaatgacttc atgttttaat tttgaccgtt 12224ctcatgttct cacataaaaa catttttcat 12284cacaacataa aatactatgt aaatcagcca 12344tataatatta ctattgatat tcactcaaat 12404ttctggttaa tatatttctt ttttaaaaca c 12464tatataattt tttcaacat ttaaccaatt 12524aattatattg agttttattt atcttttaac 12584ttattcaagt tctaagttac ttcattttta 12644tcaaaccgtt tgaagatggt ttcctcatgg 12704gctc 12738 <211> 3568 <212> DNA

<213> Lotus japonicus<220><221> 5'UTR<213> Lotus japonicus <220> <221> 5'UTR

<222> (1)..(137)<220><222> (1) .. (137) <220>

<221> CDS<222> (138)..(3119)<220><221> CDS <222> (138) .. (3119) <220>

<221> 3'UTR<222> (3120)..(3568)<400> 5<221> 3'UTR <222> (3120) .. (3568) <400> 5

ttcaactttc aaaacaaagt ggatgggatc ttcatcttat ataaccacac atcaatcatt 60tgtgctactt ctccaatttt ctttagagat gaaatgaaga gctaagcaga caagacaagt 120ttatttgttt gttgctg atg ggt ctt ggg ttc aag atg cag cag age cac 170ttcaactttc aaaacaaagt ggatgggatc ttcatcttat ataaccacac atcaatcatt 60tgtgctactt ctccaatttt ctttagagat gaaatgaaga gagtaagcaga caagacaagt 120ttatttgttt gttgctg atg ggt ctt gg cgag

Met Gly Leu Gly Phe Lys Met Gln Gln Ser His15 10Met Gly Read Gly Phe Lys Met Gln Gln Ser His15 10

cac cct gtg gct ttg aag tta cat gag caa gct ggg age cag aga aag 218His Pro vai Ala Leu Lys Leu His Glu Gln Ala Gly Ser Gln Arg Lys15 20 25cac cct gtg gtt ttg aag tta cat gag caa gct ggg age cag aga aag 218His Pro Goes Wing Leu Lys Leu His Glu Gln Wing Gly Ser Gln Arg Lys15 20 25

ttc act ttc att cag aac ttc aga aac tgg ttt cta ccc ctt ctg ttt 266Phe Thr Phe Ile Gln Asn Phe Arg Asn Trp Phe Leu Pro Leu Leu Phe30 35 40ttc act ttc att cag aac ttc aga aac tgg ttt cta ccc ctt ctg ttt 266Phe Thr Phe Ile Gln Asn Phe Arg Asn Trp Phe Leu Pro Leu Phe30 35 40

gta tgg ttc att gtt atg gct gea ttt ggt gcc tgc ate tac cat aaa 314vai Trp Phe Ile Val Met Ala Ala Phe Gly Ala Cys Ile Tyr His Lysgta tgg ttc att gtt atg gct gea ttt ggt gcc tgc till tac cat aaa 314vai Trp Phe Ile Val Met Wing Phe Gly Wing Cys Ile Tyr His Lys

45 50 5545 50 55

atg gat gct gaa act aaa gtc aga agg aaa gag gtg ctg ggt age ctc 362Met Asp Ala Glu Thr Lys Val Arg Arg Lys Glu Val Leu Gly Ser Leu60 65 70 75atg gat gct gaa act aaa gtc aga agg aaa gag gtg ctg ggt age ctc 362Met Asp Glu Thr Lys Val Arg Arg Lys Glu Val Leu Gly Ser Leu60 65 70 75

tgt gat caa agg gct aga atg cta caa gac caa ttc agt gtc agt gtc 410Cys Asp Gln Arg Ala Arg Met Leu Gln Asp Gln Phe Ser vai Ser Val80 85 90tgt gat caa agg gct aga atg cta caa gac caa ttc agt gtc agt gtc 410Cys Asp Gln Arg Wing Arg Met Leu Gln Asp Gln Phe Ser Will Be Val80 85 90

aac cat gtc cat gcc ctt gcc ate ctt gtt tca acc ttc cat tac tac 458Asn His vai His Ala Leu Ala Ile Leu vai Ser Thr Phe His Tyr Tyr95 100 105aac cat gtc cat gcc ctt gcc until ctt gtt tca acc ttc cat tac tac 458Asn His Goes His Ala Leu Ala Ile Leu Will Be Thr Phe His Tyr Tyr95 100 105

aga aat act tca gcc att gac cag gaa acc ttt gea gaa tac acg gcc 506Arg Asn Thr Ser Ala Ile Asp Gln Glu Thr Phe Ala Glu Tyr Thr Alaaga aat act tca gcc att gac cag gaa acc ttt gea gaa tac acg gcc 506Arg Asn Thr Ser Ala Ile Asp Gln Glu Thr Phe Ala Glu Tyr Thr Ala

110 115 120110 115 120

agg aca gea ttt gaa cgg cca tta atg agt ggg gtg gcc tat gea cag 554Arg Thr Ala Phe Glu Arg Pro Leu Met Ser Gly vai Ala Tyr Ala Gln125 130 135agg aca gea ttt gaa cgg cca tta atg agt ggg gtg gcc tat gea cag 554Arg Thr Ala Phe Glu Arg Pro Leu Met Ser Gly goes Ala Tyr Ala Gln125 130 135

aga gtg gtt cac tca gag aga gaa aga ttt gag aag caa cat ggg tgg 602Arg Val Val His Ser Glu Arg Glu Arg Phe Glu Lys Gln His Gly Trp140 145 150 155aga gtg gtt cac tca gag aga gaa aga ttt gag aag caa cat cat ggg tgg 602Arg Val Val His Glu Arg Glu Arg Phe Glu Lys Gln His Gly Trp140 145 150 155

gtt ata aag aca atg gaa aga gtg cct tca ggg gtt agg gat gag tat 650Val Ile Lys Thr Met Glu Arg Val Pro Ser Gly vai Arg Asp Glu Tyrgtt ata aag aca atg gaa aga gtg cct tca ggg gtt agg gat gag tat 650Val Ile Lys Thr Met Glu Arg Val Pro Ser Gly will Arg Asp Glu Tyr

160 165 170160 165 170

gea gea gtg ata ttt gea cag gaa act gtc tet tac ctt gaa tet att 698Ala Ala vai Ile Phe Ala Gln Glu Thr vai Ser Tyr Leu Glu Ser Ile175 180 185gea gea gtg ata ttt gea cag gaa act gtc tet tac ctt gaa tet att 698Ala Ala Go Ile Phe Ala Gln Glu Thr Will Be Tyr Leu Glu Ser Ile175 180 185

gat atg atg tet ggg gag gag gac cga gag aac att ttg agg gct aga 746Asp Met Met Ser Gly Glu Glu Asp Arg Glu Asn Ile Leu Arg Ala Arggat atg atg tet ggg gag gag gac cga gag aac att ttg agg gct aga 746Asp Met Met Ser Gly Glu Glu Asp Arg Glu Asn Ile Leu Arg Ala Arg

190 195 200190 195 200

gcc act ggg aaa gct gtt ctg act age cct ttc aga ctg ctg gat tet 794Ala Thr Gly Lys Ala Val Leu Thr ser Pro Phe Arg Leu Leu Asp Ser205 210 215gcc act ggg aaa gct gtt ctg act age cct ttc aga ctg ctg gat tet 794Ala Thr Gly Lys Ala Val Leu Thr be Pro Phe Arg Leu Leu Asp Ser205 210 215

cat cat ctt ggc gtg gtt cta aca ttt cct gtt tat aaa tet aag ctc 842His His Leu Gly vai vai Leu Thr Phe Pro vai Tyr Lys ser Lys Leu220 225 230 235cat cat ctt ggc gtg gtt cta aca ttt cct gtt tat aaa tet aag ctc 842His His Leu Gly Goes Goes Leu Thr Phe Pro Goes Tyr Lys Be Lys Leu220 225 230 235

cct cca gag cca acg acg gaa gag gtc att aaa gcc ata gea gga tat 890Pro Pro Glu Pro Thr Thr Glu Glu Val Ile Lys Ala lie Ala Gly Tyrcct cca gag cca acg acg gaa gag gtc att aaa gcc ata gea gga tat 890Pro Pro Glu Pro Thr Thr Glu Glu Val Ile Lys Ala lie Ala Gly Tyr

240 245 250240 245 250

att gga gga tcc ttt gat gtt gag tcc ctt gtg gag aat tta ttt ggt 938Ile Gly Gly Ser Phe Asp Val Glu Ser Leu Val Glu Asn Leu Phe Glyatt gga gga tcc ttt gat gtt gag tcc ctt gtg gag aat tta ttt ggt 938Ile Gly Gly Ser Phe Asp Val Glu Ser Leu Val Glu Asn Leu Phe Gly

255 260 265255 260 265

caa ctt gct ggt aac caa gca att ttg gtg aag gta tat gat ata aca 986Gln Leu Ala Gly Asn Gln Ala Ile Leu Val Lys Val Tyr Asp Ile Thr 270 275 280caa ctt gct ggt aac caa gca att ttg gtg aag gta tat gat ata aca 986Gln Leu Wing Gly Asn Gln Wing Ile Leu Val Lys Val Tyr Asp Ile Thr 270 275 280

aac tct age gac ccc cta ate atg tat ggc age caa tat gaa gag ggt 1034Asn Ser Ser Asp Pro Leu Ile Met Tyr Gly Ser Gln Tyr Glu Glu Glyaac tct age gac ccc cta till atg tat ggc age caa tat gaa gag ggt 1034Asn Ser Ser Asp Pro Leu Ile Met Tyr Gly Ser Gln Tyr Glu Glu Gly

285 290 295285 290 295

gat atg tct ctt gtc cat gaa agt aag ctt gat ttt gga gat cca tac 1082Asp Met Ser Leu Val His Glu Ser Lys Leu Asp Phe Gly Asp Pro Tyr300 305 310 315gat atg tct ct gtc cat gaa agt aag ctt gat ttt gga gat cca tac 1082Asp Met Ser Leu Val His Glu Ser Lys Leu Asp Phe Gly Asp Pro Tyr300 305 310 315

agg aaa cat cac atg ate tgt aga tat cac caa cag gca cca aca aat 1130Arg Lys His His Met Ile Cys Arg Tyr His Gln Gln Ala Pro Thr Asn320 325 330agg aaa cat cac atg till tgt aga tat cac caa cag gca cca aca aat 1130Arg Lys His His Met Ile Cys Arg Tyr His Gln Ala Pro Thr Asn320 325 330

tgg ata gca tat acc acg gca ttc cta ttc ttt gtg att ctt tgt tta 1178Trp Ile Ala Tyr Thr Thr Ala Phe Leu Phe Phe Val Ile Leu Cys Leutgg ata gca tat acc acg gca ttc cta ttc ttt gtg att ctt tgt tta 1178Trp Ile Wing Tyr Thr Thr Wing Phe Leu Phe Val Ile Leu Cys Leu

335 340 345335 340 345

gtg ggt tac att tta tat gct gct gga act cac att gtc aag gta gaa 1226Val Gly Tyr Ile Leu Tyr Ala Ala Gly Thr His Ile vai Lys Val Glu 350 355 360gtg ggt tac att tta tat gct gct gga act cac att gtc aag gta gaa 1226Val Gly Tyr Ile Leu Tyr Wing Gly Thr His Ile Goes Lys Val Glu 350 355 360

gat gat tac aat gca atg cag gat tta aaa gtc aaa gca gaa gca gct 1274Asp Asp Tyr Asn Ala Met Gln Asp Leu Lys vai Lys Ala Glu Ala Alagat gat tac aat gca atg cag gat tta aaa gtc aaa gca gaa gca gct 1274Asp Asp Tyr Asn Ala Met Gln Asp Leu Lys Goes Lys Ala Glu Ala Ala

365 370 375365 370 375

gat att gcc aag tca cag ttt cta gct acc gtc tct cat gaa att aga 1322Asp Ile Ala Lys Ser Gln Phe Leu Ala Thr vai Ser His Glu Ile Arg380 385 390 395gat att gcc aag tca cag ttt cta gct acc gtc tct cat gaa att aga 1322Asp Ile Wing Lys Ser Gln Phe Wing Wing Thr Will Be His Glu Ile Arg380 385 390 395

act ccc atg aat gga att tta gga atg ctt ggt ctg ctt tta cgc aca 1370Thr Pro Met Asn Gly Ile Leu Gly Met Leu Gly Leu Leu Leu Arg Thr400 405 410act ccc atg aat gga att tta gga atg ctt ggt ctg ctt tta cgc aca 1370Thr Pro Met Asn Gly Ile Leu Gly Met Leu Gly Leu Leu Arg Thr400 405 410

gaa ttg agt tca aca caa aga gac tat gct cag act gct caa gca tgt 1418Glu Leu Ser Ser Thr Gln Arg Asp Tyr Ala Gln Thr Ala Gln Ala Cys415 420 425ggg aag gca cta ata gca tta ata aat gag gtg ctt gac cga gct aaa 1466Gly Lys Ala Leu lie Ala Leu lie Asn Glu Val Leu Asp Arg Ala Lysgaa ttg agt tca aca caa aga gac tat gct cag act gct caa gca tgt 1418Glu Leu Being Ser Thr Gln Arg Asp Tyr Ala Gln Thr Ala Gln Ala Cys415 4204 425ggg aag gca cta ata gca gta ctt gac cga gly cga 14 Lys Wing Leu lie Wing Leu lie Asn Glu Val Leu Asp Arg Wing Lys

430 435 440430 435 440

att gaa gca ggc aaa tta gag cta gaa gca gtt cca ttt gac ctt cgt 1514att gaa gca ggc aaa tta gag cta gaa gca gtt cca ttt gac ctt cgt 1514

Ile Glu Ala Gly Lys Leu Glu Leu Glu Ala Val Pro Phe Asp Leu Arg445 450 455Ile Glu Wing Gly Lys Leu Glu Leu Glu Leu Val Pro Phe Asp Leu Arg445 450 455

tcc ata ctt gac gat gtc ctt tct ctt ttt tct gag aag tca aga cac 1562ser Ile Leu Asp Asp vai Leu Ser Leu Phe Ser Glu Lys Ser Arg His460 465 470 475tcc ata ctt gac gat gtc ctt tct ctt ttt tct gag aag tca aga cac 1562ser Ile Leu Asp Asp will Leu Ser Leu Phe Ser Glu Lys Ser Arg His460 465 470 475

aaa ggc tta gag ctg gca gtg ttt gtt tct gac aaa gtt ccg gat ata 1610Lys Gly Leu Glu Leu Ala Val Phe Val Ser Asp Lys Val Pro Asp Ileaaa ggc tta gag ctg gca gtg ttt gtt tct gac aaa gtt ccg gat ata 1610Lys Gly Leu Glu Leu Ala Val Phe Val Ser Asp Lys Val Pro Asp Ile

480 485 490480 485 490

gtt atg ggc gat cct ggg cga ttc aga caa ata gtg aca aat ctt gtt 1658vai Met Gly Asp Pro Gly Arg Phe Arg Gln Ile vai Thr Asn Leu Val495 500 505gt atg ggc gat cct ggg cga ttc aga caa ata gtg aca aat ctt gtt 1658vai Met Gly Asp Pro Gly Arg Phe Arg Gln Ile Go Thr Asn Leu Val495 500 505

gga aac tct gtt aag ttc act gag cga ggt cat ata ttt gtt aaa gtc 1706Gly Asn Ser vai Lys Phe Thr Glu Arg Gly His Ile Phe vai Lys vaigga aac tct gtt aag ttc act gag cga ggt cat ata ttt gtt aaa gtc 1706Gly Asn Ser Will Lys Phe Thr Glu Arg Gly His Ile Phe Will Lys Will

510 515 520510 515 520

cat tta gct gaa aaa aga cag tgc aca atg aat gga aaa tgt gag act 1754His Leu Ala Glu Lys Arg Gln Cys Thr Met Asn Gly Lys Cys Glu Thr525 530 535cat tta gct gaa aaa aga cag tgc aca atg aat gga aaa tgt gag act 1754His Leu Wing Glu Lys Arg Gln Cys Thr Met Asn Gly Lys Cys Glu Thr525 530 535

ttt cta aat gga ggc tgt gat gat gtt ttg cat gta tct ggc agt tat 1802Phe Leu Asn Gly Gly Cys Asp Asp Val Leu His Val Ser Gly Ser Tyr540 545 550 555ttt cta aat gga ggc tgt gat gat gtt ttg cat gta tct ggc agt tat 1802Phe Leu Asn Gly Gly Cys Asp Asp Val Leu His Val Ser Gly Ser Tyr540 545 550 555

aat ttg aaa acc ctt agt gga tat gaa gcc gct gat gaa cgg aac age 1850Asn Leu Lys Thr Leu Ser Gly Tyr Glu Ala Ala Asp Glu Arg Asn Seraat ttg aaa acc ctt agt gga tat gaa gcc gct gat gaa cgg aac age 1850Asn Leu Lys Thr Leu Ser Gly Tyr Glu Ala Wing Asp Glu Arg Asn Ser

560 565 570560 565 570

tgg gat aat ttt aag cat cat att gct gac gaa gaa ttt ttc ttt gat 1898Trp Asp Asn Phe Lys His His Ile Ala Asp Glu Glu Phe Phe Phe Asp575 580 585tgg gat aat ttt aag cat cat att gct gac gaa gaa ttt ttc ttt gat 1898Trp Asp Asn Phe Lys His His Ile Ala Asp Glu Glu Phe Phe Phe Asp575 580 585

gct tcg gtt aaa aag ttg gcc tct agt gaa tct tat gag caa gtc acc 1946Ala Ser Val Lys Lys Leu Ala Ser Ser Glu Ser Tyr Glu Gln Val Thr590 595 600gct tcg gtt aaa aag ttg gcc tct agt gaa tct tat gag caa gtc acc 1946Ala Ser Val Lys Lys Leu Ala Ser Ser Glu Ser Tyr Gln Val Thr590 595 600

ttg atg gtc age gtg gag gac act gga att ggg att tet ttc tet gcc 1994Leu Met vai Ser Val Glu Asp Thr Gly Ile Gly Ile Ser Phe Ser Ala605 610 615ttg atg gtc age gtg gag gac act gga att ggg att tet ttc tet gcc 1994Leu Met Will Be Val Glu Asp Thr Gly Ile Gly Ile Ser Phe Ser Ala605 610 615

caa gat agt att ttc atg cct ttt gtg cag gct gac age tca acc tet 2042Gln Asp Ser Ile Phe Met Pro Phe vai Gln Ala Asp Ser Ser Thr Ser620 625 630 635caa gat agt att ttc atg cct ttt gtg cag gct gac age tca acc tet 2042Gln Asp Ser Ile Phe Met Pro Phe Go Gln Wing Asp Ser Ser Thr Ser620 625 630 635

cga aac tat ggg ggt acc ggg ate ggc ttg agt ate agt aag tgc ttg 2090Arg Asn Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile Ser Lys Cys Leu640 645 650cga aac tat ggg ggt acc ggg to ggc ttg agt to agt aag tgc ttg 2090Arg Asn Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile Ser Lys Cys Leu640 645 650

gtt gaa ctg atg ggc ggt cag ata aac ttc ata age cga ccc cag gtc 2138vai Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Ser Arg Pro Gln vai655 660 665gaa ctg atg gtt cag ggc ggt ttc aac ata ata act cga gtc ccc cag 2138vai Leu Glu Met Gly Gly Ile Gln Asn Phe Ser Arg Ile Pro Gln 660 665 vai655

ggg age acg ttt tca ttc act gea gat ttc gga aca ttt aag aaa aac 218615 Gly Ser Thr Phe Ser Phe Thr Ala Asp Phe Gly Thr Phe Lys Lys Asn670 675 680ggg age acg ttt tca ttc act gea gat ttc gga aca ttt aag aaa aac 218615 Gly Ser Thr Phe Ser Phe Thr Asp Phe Gly Thr Phe Lys Lys Asn670 675 680

tca aca act gac atg aag aaa ctt aac ttt gaa gat cta cct tet agt 2234Ser Thr Thr Asp Met Lys Lys Leu Asn Phe Glu Asp Leu Pro Ser Ser685 690 695tca aca act gac atg aag aaa ctt aac ttt gaa gat cta cct tet agt 2234Ser Thr Thr Asp Met Lys Lys Leu Asn Phe Glu Asp Leu Pro Ser685 690 695

ttt aga ggt ctt aaa gcc att gtg gtt gat gga aaa cct gtt aga gct 2282Phe Arg Gly Leu Lys Ala Ile Val Val Asp Gly Lys Pro Val Arg Ala700 705 710 715ttt aga ggt ctt aaa gcc att gtg gtt gat gga aaa cct gtt aga gct 2282Phe Arg Gly Leu Lys Ala Ile Val Val Asp Gly Lys Pro Val Arg Ala700 705 710 715

gea gtg act aga tac cat ttg aag aga cta ggg ata caa gct aaa gtt 2330Ala vai Thr Arg Tyr His Leu Lys Arg Leu Gly Ile Gln Ala Lys vai720 725 730gea gtg act aga tac cat ttg aag aga cta ggg ata caa gct aaa gtt 2330Ala goes Thr Arg Tyr His Leu Lys Arg Leu Gly Ile Gln Ala Lys vai720 725 730

gea att age ate aat aag gct gtt tet tta tgt ggg aaa aat ggt tet 2378Ala Ile Ser Ile Asn Lys Ala vai Ser Leu Cys Gly Lys Asn Gly Ser735 740 745gea att age till aat aag gct gtt tet tta tgt ggg aaa aat ggt tet 2378Ala Ile Ser Ile Asn Lys Ala Will Be Leu Cys Gly Lys Asn Gly Ser735 740 745

ttg acc tcg gea tta ttt cag cct gat att att ttt gtt gag aag gac 2426Leu Thr Ser Ala Leu Phe Gln Pro Asp Ile Ile Phe Val Glu Lys Asp750 755 760ttg acc tcg gea tta ttt cag cct gat att att ttt gtt gag aag gac 2426Leu Thr Be Wing Leu Phe Gln Pro Asp Ile Ile Phe Val Glu Lys Asp750 755 760

tet tgg gtt tet gga gag gat ggt ggt ate ttc aat gcg ttt aag atg 2474Ser Trp Val Ser Gly Glu Asp Gly Gly Ile Phe Asn Ala Phe Lys Mettet tgg gtt tet gga gag gat ggt ggt until ttc aat gcg ttt aag atg 2474Ser Trp Val Ser Gly Glu Asp Gly Ile Phe Asn Ala Phe Lys Met

765 770 775765 770 775

cct caa atg ate ctt ctt gea acc aat ate tgt aac gct gaa ttt gat 2522Pro Gln Met Ile Leu Leu Ala Thr Asn Ile Cys Asn Ala Glu Phe Asp780 785 790 795cct caa atg till ctt ctt gea acc aat till tgt aac gct gaa ttt gat 2522Pro Gln Met Ile Leu Leu Thr Wing Asn Ile Cys Asn Wing Glu Phe Asp780 785 790 795

aaa gcc aaa gct gea ggt ttc agt gat aca gtg ate atg aag cca ctg 2570Lys Ala Lys Ala Ala Gly Phe Ser Asp Thr vai Ile Met Lys Pro Leuaaa gcc aaa gct gea ggt ttc agt gat aca gtg till atg aag cca ctg 2570Lys Ala Lys Ala Ala Gly Phe Ser Asp Thr Go Ile Met Lys Pro Leu

800 805 810800 805 810

aga gct agt atg ctg gct gct tgt ctt cag caa gtt ttc ggg act ggc 2618Arg Ala Ser Met Leu Ala Ala Cys Leu Gln Gln Val Phe Gly Thr Gly815 820 825aga gct agt atg ctg gct gct tgt ctt cag caa gtt ttc ggg act ggc 2618Arg Ala Ser Met Leu Ala Cys Leu Gln Gln Val Phe Gly Thr Gly815 820 825

aag acg agg cag ttt ggg aaa gac atg tcg aat ggt tet tca gta cga 2666Lys Thr Arg Gln Phe Gly Lys Asp Met Ser Asn Gly Ser Ser Val Arg830 835 840aag acg agg cag ttt ggg aaa gac atg tcg aat ggt tet tca gta cga 2666Lys Thr Arg Gln Phe Gly Lys Asp Met Ser Asn Gly Ser Ser Val Arg830 835 840

age ctt ctt tgc gga aag aaa ate tta gtg gtt gat gat aat ttg gtg 2714Ser Leu Leu Cys Gly Lys Lys Ile Leu Val Val Asp Asp Asn Leu Valage ctt ctt tgc gga aag aaa till tta gtg gtt gat gat aat ttg gtg 2714Ser Leu Leu Cys Gly Lys Lys Ile Leu Val Val Asp Asp Asn Leu Val

845 850 855845 850 855

aac cga agg gtc gcc gcc ggc gcg ttg aaa aac ttt gga gct gat gtc 2762Asn Arg Arg Val Ala Ala Gly Ala Leu Lys Asn Phe Gly Ala Asp Val860 865 870 875aac cga agg gtc gcc gcc ggc gcg ttg aaa aac ttt gga gct gat gtc 2762Asn Arg Arg Val Wing Wing Gly Wing Leu Lys Asn Phe Gly Wing Wing Val860 865 870 875

aaa tgt gea gea agt ggc aaa gct gct ctt gaa atg ctt caa tat cct 2810Lys Cys Ala Ala Ser Gly Lys Ala Ala Leu Glu Met Leu Gln Tyr Proaaa tgt gea gea agt ggc aaa gct gct ctt gaa atg ctt caa tat cct 2810Lys Cys Wing Wing Ser Gly Lys Wing Wing Leu Glu Met Leu Gln Tyr Pro

880 885 890880 885 890

cac gat ttc gat gct tgc ttc atg gat att caa atg cca gaa atg gat 2858His Asp Phe Asp Ala Cys Phe Met Asp Ile Gln Met Pro Glu Met Asp895 900 905cac gat ttc gat gct tgc ttc atg gat att caa atg cca gaa atg gat 2858His Asp Phe Asp Cys Phe Met Asp Ile Gln Met Pro Glu Met Asp895 900 905

ggg ttt gag gcc act cgt cga att cgg atg atg gaa aga gag gea agt 2906Gly Phe Glu Ala Thr Arg Arg Ile Arg Met Met Glu Arg Glu Ala Ser910 915 920ggg ttt gag gcc act cgt cga att cgg atg atg gaa aga gag gea agt 2906Gly Phe Glu Ala Thr Arg Arg Ile Arg Met Met Glu Arg Glu Ala Ser910 915 920

gag cag ctg aaa agt gaa tet ggt gaa gaa aat ggt aag aaa agt gag 2954Glu Gln Leu Lys Ser Glu Ser Gly Glu Glu Asn Gly Lys Lys Ser Glu925 930 935ttc cac atg cct ata ttg gcc atg aca gct gat gta ate cat gct aca 3002Phe His Met Pro Ile Leu Ala Met Thr Ala Asp Val Ile His Ala Thr940 945 950 955gag cag ctg aaa agt gaa tet ggt gaa gaa aat ggt aag aaa agt gag 2954Glu Gln Leu Lys Be Glu Ser Gly Glu Asn Gly Lys Lys Ser Glu925 930 935ttc cac atg cct ata ttg gcc atg acc gct gat gta 300 His Met Pro Ile Read Wing Met Thr Wing Asp Val Ile His Wing Thr940 945 950 955

tat gat aag tgc tta aat tgt ggg atg gat gga tac gtc tca aag cct 3050Tyr Asp Lys Cys Leu Asn Cys Gly Met Asp Gly Tyr vai ser Lys Protat gat aag tgc tta aat tgt ggg atg gat gga tac gtc tca aag cct 3050Tyr Asp Lys Cys Leu Asn Cys Gly Met Asp Gly Tyr is going to be Lys Pro

960 965 970960 965 970

ttt gaa gaa gag aat ctc tat caa gea gtt gea aag ttc ttc aag tcc 3098Phe Glu Glu Glu Asn Leu Tyr Gln Ala vai Ala Lys Phe Phe Lys Ser975 980 985ttt gaa gaa gag aat ctc tat caa gea gtt gea aag ttc ttc aag tcc 3098Phe Glu Glu Glu Asn Leu Tyr Gln

aag cct gct tca gac tca tga cactgcttat tctgcagaac aggtcaacca 3149aag cct gct tca gac tca tga cactgcttat tctgcagaac aggtcaacca 3149

Lys Pro Ala Ser Asp Ser990Lys Pro Wing Ser Asp Ser990

acttttgatt gagaaacatt tagtgttagc atgtttggat caacttctcc cagcatcaat 3209tctgaaactc agaagctact catagaagct tctagccaga attgattttg gcttcagaag 3269ctactaatgg ttttatgtag agagcaaaag ttggtttcca agatatgcga ggatccatga 3329tgatacacac gtctgaagtg ateattteta accagttgaa gtttcactcg acgtgatttg 3389aatccaagta aatgcatacc acataattat ccatccccat cttgtgtaca gattctccca 3449agggataaga aaatttatgt aaattcaatt tttttctttt gcatctcaat acttccctgt 3509tagaactttt tccctatgat tatttccact tttcattttc aattatattt tttgtaaaa 3568<210> 6<211> 993<212> PRTacttttgatt gagaaacatt tagtgttagc atgtttggat caacttctcc cagcatcaat 3209tctgaaactc agaagctact catagaagct tctagccaga attgattttg gcttcagaag 3269ctactaatgg ttttatgtag agagcaaaag ttggtttcca agatatgcga ggatccatga 3329tgatacacac gtctgaagtg ateattteta accagttgaa gtttcactcg acgtgatttg 3389aatccaagta aatgcatacc acataattat ccatccccat cttgtgtaca gattctccca 3449agggataaga aaatttatgt aaattcaatt tttttctttt gcatctcaat acttccctgt 3509tagaactttt tccctatgat tatttccact tttcattttc aattatattt tttgtaaaa 3568 <210> 6 <211> 993 <212> PRT

<213> Lotus japonicus<400> 6<213> Lotus japonicus <400> 6

Met Gly Leu Gly Phe Lys Met Gln Gln ser His His Pro vai Ala Leu1 5 10 15Met Gly Leu Gly Phe Lys Met Gln Gln Be His His Pro Goes Wing Leu1 5 10 15

Lys Leu His Glu Gln Ala Gly Ser Gln Arg Lys Phe Thr Phe Ile GlnLys Read His Glu Gln Wing Gly Be Gln Arg Lys Phe Thr Phe Ile Gln

20 25 3020 25 30

Asn Phe Arg Asn Trp Phe Leu Pro Leu Leu Phe vai Trp Phe Il e vai 35 40 45Asn Phe Arg Asn Trp Phe Leu Pro Leu Leu Phe Goes Trp Phe Il And Goes 35 40 45

Met Ala Ala Phe Gly Ala Cys Ile Tyr His Lys Met Asp Ala Glu Thr50 55 60Lys Val Arg Arg Lys Glu Val Leu Gly Ser Leu Cys Asp Gln Arg Ala65 70 75 80Met Wing Phe Wing Gly Cys Wing Ile Tyr His Lys Met Asp Wing Glu Thr50 55 60Lys Val Arg Arg Lys Glu Val Leu Gly Ser Leu Cys Asp Gln Arg Ala65 70 75 80

Arg Met Leu Gln Asp Gln Phe Ser Val Ser Val Asn His Val His Ala85 90 95Arg Met Read Gln Asp Gln Phe Ser Val Ser Val Asn His Val His Ala85 90 95

Leu Ala Ile Leu Val Ser Thr Phe His Tyr Tyr Arg Asn Thr ser Ala100 105 110Leu Ala Ile Leu Val Be Thr Phe His Tyr Tyr Arg Asn Thr Be Ala100 105 110

Ile Asp Gln Glu Thr Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe GluIle Asp Gln Glu Thr Phe Wing Glu Tyr Glou Arg Wing Thr Phe Glu

115 120 125115 120 125

Arg Pro Leu Met Ser Gly Val Ala Tyr Ala Gln Arg Val Val His Ser130 135 140Arg Pro Read Met Ser Gly Val Wing Tyr Wing Gln Arg Val Val His Ser130 135 140

Glu Arg Glu Arg Phe Glu Lys Gln His Gly Trp vai Ile Lys Thr Met145 150 155 160Glu Arg Glu Arg Phe Glu Lys Gln His Gly Trp Goes Ile Lys Thr Met145 150 155 160

Glu Arg Val Pro Ser Gly vai Arg Asp Glu Tyr Ala Ala Val Ile Phe165 170 175Glu Arg Val Pro Being Gly Goes Arg Asp Glu Tyr Wing Val Ile Wing Phe165 170 175

Ala Gln Glu Thr vai ser Tyr Leu Glu Ser Ile Asp Met Met Ser Gly180 185 190Gln Glu Wing Thr will be Tyr Leu Glu Ser Ile Asp Met Met Ser Gly180 185 190

Glu Glu Asp Arg Glu Asn Ile Leu Arg Ala Arg Ala Thr Gly Lys AlaGlu Glu Asp Arg Glu Asn Ile Read Leu Arg Wing Arg Wing Thr Gly Lys Wing

195 200 205195 200 205

Val Leu Thr Ser Pro Phe Arg Leu Leu Asp Ser His His Leu Gly Val210 215 220Val Leu Thr Be Pro Phe Arg Leu Read Asp Be His His Leu Gly Val210 215 220

Val Leu Thr Phe Pro vai Tyr Lys Ser Lys Leu Pro Pro Glu Pro Thr225 230 235 240Val Leu Thr Phe Pro will Tyr Lys Be Lys Leu Pro Pro Glu Pro Thr225 230 235 240

Thr Glu Glu vai Ile Lys Ala Ile Ala Gly Tyr Ile Gly Gly Ser Phe245 250 255Thr Glu Glu goes Ile Lys Wing Ile Wing Gly Tyr Ile Gly Gly Ser Phe245 250 255

Asp vai Glu Ser Leu vai Glu Asn Leu Phe Gly Gln Leu Ala Gly Asn260 265 270Asp goes Glu Be Leu goes Glu Asn Leu Phe Gly Gln Leu Wing Gly Asn260 265 270

Gln Ala Ile Leu Val Lys Val Tyr Asp Ile Thr Asn Ser Ser Asp ProGln Wing Ile Read Val Lys Val Tyr Asp Ile Thr Asn Be Ser Asp Pro

275 280 285275 280 285

Leu Ile Met Tyr Gly Ser Gln Tyr Glu Glu Gly Asp Met Ser Leu Val290 295 300Leu Ile Met Tyr Gly Ser Gln Tyr Glu Glu Gly Asp Met Ser Leu Val290 295 300

His Glu Ser Lys Leu Asp Phe Gly Asp Pro Tyr Arg Lys His His Met305 310 315 320Ile Cys Arg Tyr His Gln Gln Ala Pro Thr Asn Trp Ile Ala Tyr ThrHis Glu Ser Lys Read Asp Phe Gly Asp Pro Tyr Arg Lys His His Met305 310 315 320Ile Cys Arg Tyr His Gln Gln Wing Pro Thr Asn Trp Ile Wing Tyr Thr

325 330 335325 330 335

Thr Ala Phe Leu Phe Phe Val lie Leu Cys Leu Val Gly Tyr Ile Leu340 345 350Thr Ala Phe Leu Phe Phe Val lie Leu Cys Leu Val Gly Tyr Ile Leu340 345 350

Tyr Ala Ala Gly Thr His Ile vai Lys Val Glu Asp Asp Tyr Asn Ala355 360 365Tyr Wing Ally Gly Thr His Ile Goes Lys Val Glu Asp Asp Tyr Asn Ala355 360 365

Met Gln Asp Leu Lys Val Lys Ala Glu Ala Ala Asp Ile Ala Lys SerMet Gln Asp Leu Lys Val Lys Wing Glu Wing Asp Wing Ile Wing Lys Ser

370 375 380370 375 380

Gln Phe Leu Ala Thr vai Ser His Glu Ile Arg Thr Pro Met Asn Gly 385 390 395 400Gln Phe Leu Wing Thr Will Be His Glu Ile Arg Thr Pro Met Asn Gly 385 390 395 400

lie Leu Gly Met Leu Gly Leu Leu Leu Arg Thr Glu Leu Ser Ser Thrlie Leu Gly Met Leu Gly Leu Leu Leu Arg Thr Glu Leu Being Ser Thr

405 410 415405 410 415

Gln Arg Asp Tyr Ala Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu Ile420 425 430Gln Arg Asp Tyr Wing Gln Thr Wing Gln Cys Wing Gly Lys Wing Leu Ile420 425 430

Ala Leu Ile Asn Glu Val Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys435 440 445Wing Leu Ile Asn Glu Val Leu Asp Arg Wing Lys Ile Glu Wing Gly Lys435 440 445

Leu Glu Leu Glu Ala Val Pro Phe Asp Leu Arg Ser Ile Leu Asp AspLeu Glu Leu Glu Val Wing Val Pro Phe Asp Leu Arg Ser Ile Leu Asp Asp

450 455 460450 455 460

vai Leu Ser Leu Phe Ser Glu Lys Ser Arg His Lys Gly Leu Glu Leu 465 470 475 480will Read Be Read Be Phe Be Glu Lys Be Arg His Lys Gly Leu Glu Leu 465 470 475 480

Ala Val Phe vai Ser Asp Lys vai Pro Asp Ile Val Met Gly Asp ProVal Phe Wing Will Be Asp Lys Go Pro Asp Ile Val Met Gly Asp Pro

485 490 495485 490 495

Gly Arg Phe Arg Gln Ile Val Thr Asn Leu Val Gly Asn Ser Val Lys500 505 510Gly Arg Phe Arg Gln Ile Val Thr Asn Read Val Gly Asn Ser Val Lys500 505 510

Phe Thr Glu Arg Gly His Ile Phe Val Lys Val His Leu Ala Glu Lys515 520 525Phe Thr Glu Arg Gly His Ile Phe Val Lys Val His Leu Wing Glu Lys515 520 525

Arg Gln Cys Thr Met Asn Gly Lys Cys Glu Thr Phe Leu Asn Gly GlyArg Gln Cys Thr Met Asn Gly Lys Cys Glu Thr Phe Read Asn Gly Gly

530 535 540530 535 540

Cys Asp Asp vai Leu His vai Ser Gly Ser Tyr Asn Leu Lys Thr Leu 545 550 555 560Cys Asp Asp Will Leu His Will Be Gly Ser Tyr Asn Leu Lys Thr Leu 545 550 555 560

Ser Gly Tyr Glu Ala Ala Asp Glu Arg Asn Ser Trp Asp Asn Phe Lys565 570 575His His Ile Ala Asp Glu Glu Phe Phe Phe Asp Ala Ser Val Lys LysSer Gly Tyr Glu Ala Wing Asp Glu Arg Asn Ser Trp Asp Asn Phe Lys565 570 575His His Ile Wing Asp Glu Glu Phe Phe Phe Asp Wing Ser Val Lys Lys

580 585 590580 585 590

Leu Ala Ser Ser Glu Ser Tyr Glu Gln Val Thr Leu Met Val Ser Val595 600 605Leu Wing Be Being Glu Be Tyr Glu Gln Val Thr Read Leu Met Val Be Val595 600 605

Glu Asp Thr Gly lie Gly Ile Ser Phe Ser Ala Gln Asp Ser Ile Phe610 615 620Glu Asp Thr Gly lie Gly Ile Be Phe Be Wing Gln Asp Be Ile Phe610 615 620

Met Pro Phe Val Gln Ala Asp Ser Ser Thr Ser Arg Asn Tyr Gly Gly625 630 635 640Met Pro Phe Val Gln Wing Asp Be Be Thr Be Arg Asn Tyr Gly Gly625 630 635 640

Thr Gly Ile Gly Leu Ser Ile Ser Lys Cys Leu Val Glu Leu Met Gly 645 650 655Thr Gly Ile Gly Leu Ile Be Lys Cys Leu Val Glu Leu Met Gly 645 650 655

Gly Gln Ile Asn Phe Ile Ser Arg Pro Gln vai Gly Ser Thr Phe SerGly Gln Ile Asn Phe Ile Be Arg Pro Gln Go Gly Be Thr Phe Be

660 665 670660 665 670

Phe Thr Ala Asp Phe Gly Thr Phe Lys Lys Asn Ser Thr Thr Asp Met675 680 685Phe Thr Wing Asp Phe Gly Thr Phe Lys Lys Asn Be Thr Thr Asp Met675 680 685

Lys Lys Leu Asn Phe Glu Asp Leu Pro Ser Ser Phe Arg Gly Leu Lys690 695 700Lys Lys Read Asn Phe Glu Asp Read Pro Be Ser Phe Arg Gly Leu Lys690 695 700

Ala Ile vai vai Asp Gly Lys Pro Val Arg Ala Ala vai Thr Arg Tyr705 710 715 720Wing Ile Goes Go Asp Gly Lys Pro Val Arg Wing Wing Goes Thr Arg Tyr705 710 715 720

His Leu Lys Arg Leu Gly Ile Gln Ala Lys vai Ala Ile Ser Ile Asn 725 730 735His Leu Lys Arg Leu Gly Ile Gln Wing Lys Go Wing Ile Ser Ile Asn 725 730 735

Lys Ala Val Ser Leu Cys Gly Lys Asn Gly Ser Leu Thr Ser Ala LeuLys Ala Val Be Leu Cys Gly Lys Asn Gly Be Leu Thr Be Ala Leu

740 745 750740 745 750

Phe Gln Pro Asp Ile Ile Phe Val Glu Lys Asp Ser Trp Val Ser Gly755 760 765Phe Gln Pro Asp Ile Ile Phe Val Glu Lys Asp Ser Trp Val Ser Gly755 760 765

Glu Asp Gly Gly Ile Phe Asn Ala Phe Lys Met Pro Gln Met Ile Leu770 775 780Glu Asp Gly Gly Ile Phe Asn Phe Lys Wing Pro Pro Gln Met Ile Leu770 775 780

Leu Ala Thr Asn Ile Cys Asn Ala Glu Phe Asp Lys Ala Lys Ala Ala785 790 795 800Leu Wing Thr Asn Ile Cys Asn Wing Glu Phe Asp Lys Wing Lys Wing Wing785 790 795 800

Gly Phe Ser Asp Thr Val Ile Met Lys Pro Leu Arg Ala Ser Met Leu 805 810 815Gly Phe Ser Asp Thr Val Ile Met Lys Pro Read Arg Wing Be Met Leu 805 810 815

Ala Ala Cys Leu Gln Gln Val Phe Gly Thr Gly Lys Thr Arg Gln Phe820 825 830Gly Lys Asp Met Ser Asn Gly Ser Ser Val Arg Ser Leu Leu Cys GlyWing Cys Wing Read Gln Gln Val Phe Gly Thr Gly Lys Thr Arg Gln Phe820 825 830Gly Lys Asp Met Be Asn Gly Be Be Val Arg Be Read Leu Cys Gly

835 840 845835 840 845

Lys Lys Ile Leu Val Val Asp Asp Asn Leu vai Asn Arg Arg Val Ala850 855 860Lys Lys Ile Leu Val Val Asp Asp Asn Leu Goes Asn Arg Arg Val Ala850 855 860

Ala Gly Ala Leu Lys Asn Phe Gly Ala Asp vai Lys Cys Ala Ala Ser865 870 875 880Wing Gly Wing Read Lys Asn Phe Gly Wing Asp Goes Lys Cys Wing Ser865 870 875 880

Gly Lys Ala Ala Leu Glu Met Leu Gln Tyr Pro His Asp Phe Asp AlaGly Lys Wing Wing Read Glu Met Read Gln Tyr Pro His Asp Phe Asp Wing

885 890 895885 890 895

Cys Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala Thr900 905 910Cys Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Wing Thr900 905 910

Arg Arg Ile Arg Met Met Glu Arg Glu Ala Ser Glu Gln Leu Lys SerArg Arg Ile Arg Met Met Glu Arg Glu Wing Be Glu Gln Leu Lys Ser

915 920 925915 920 925

Glu Ser Gly Glu Glu Asn Gly Lys Lys Ser Glu Phe His Met Pro Ile930 935 940Glu Being Gly Glu Glu Asn Gly Lys Lys Being Glu Phe His Met Pro Ile930 935 940

Leu Ala Met Thr Ala Asp Val Ile His Ala Thr Tyr Asp Lys Cys Leu945 950 955 960Leu Wing Met Thr Wing Asp Val Ile His Wing Thr Tyr Asp Lys Cys Leu945 950 955 960

Asn Cys Gly Met Asp Gly Tyr Val Ser Lys Pro Phe Glu Glu Glu AsnAsn Cys Gly Met Asp Gly Tyr Val Ser Lys Pro Phe Glu Glu Glu Asn

965 970 975965 970 975

Leu Tyr Gln Ala vai Ala Lys Phe Phe Lys Ser Lys Pro Ala Ser Asp980 985 990Read Tyr Gln Wing Go Wing Lys Phe Phe Lys Ser Lys Pro Wing Ser Asp980 985 990

SerTo be

<210> 7<210> 7

<211> 4743<211> 4743

<212> DNA<212> DNA

<213> Medicago truncatula<220><213> Medicago truncatula <220>

<221> exon<221> exon

<222> (1)..(348)<220><222> (1) .. (348) <220>

<221> Intron<221> Intron

<222> (349)..(462)<220><221> exon<222> (463)..(696)<220><221> Intron<222> (697)..(799)<220><221> exon<222> (800)..(969)<220><221> Intron<222> (970)..(1261)<220><221> exon<222> (1262)..(1480)<223> mutated codon encoding L/F substituti<220><221> Intron<222> (1481)..(1574)<220><221> exon<222> (1575)..(1761)<220><221> Intron<222> (1762)..(1857)<220><221> exon<222> (1858)..(1909)<220><221> intron<222> (1910)..(2018)<220><221> exon<222> (349) .. (462) <220> <221> exon <222> (463) .. (696) <220> <221> Intron <222> (697) .. (799) <220> <221> exon <222> (800) .. (969) <220> <221> Intron <222> (970) .. (1261) <220> <221> exon <222> (1262) .. (1480 ) <223> mutated codon encoding replacement L / F <220> <221> Intron <222> (1481) .. (1574) <220> <221> exon <222> (1575) .. (1761) <220> <221> Intron <222> (1762) .. (1857) <220> <221> exon <222> (1858) .. (1909) <220> <221> intron <222> (1910) .. (2018 ) <220> <221> exon

<222> (2019)..(2248)<220><222> (2019) .. (2248) <220>

<221> Intron<221> Intron

<222> (2249)..(2383)<220><222> (2249) .. (2383) <220>

<221> exon<221> exon

<222> (2384)..(2482)<220><222> (2384) .. (2482) <220>

<221> Intron<221> Intron

<222> (2483)..(2624)<220><222> (2483) .. (2624) <220>

<221> exon<221> exon

<222> (2625)..(3340)<220><222> (2625) .. (3340) <220>

<221> Intron<221> Intron

<222> (3341)..(3597)<222> (3341) .. (3597)

<220><220>

<221> exon<221> exon

<222> (3598)..(4101)<220><222> (3598) .. (4101) <220>

<221> Intron<221> Intron

<222> (4102)..(4490)<220><222> (4102) .. (4490) <220>

<221> exon<221> exon

<222> (4491)..(4743)<222> (4491) .. (4743)

<400> 7<400> 7

atg ggt ctt ctc ttg aag atg aag atg cag aat cag cac cac cct ttg 48atg ggt ctt ctc ttg aag atg aag atg cag aat cag cac cac cct ttg 48

Met Gly Leu Leu Leu Lys Met Lys Met Gln Asn Gln His His Pro LeuMet Gly Leu Leu Leu Lys Met Lys Met Gln Asn Gln His His Pro Leu

1 5 10 151 5 10 15

gct tct aag tta caa gaa caa acg ggg aac aaa aga tac aca ttc att 96gct tct aag tta caa gaa caa acg ggg aac aaa aga tac aca ttc att 96

Ala Ser Lys Leu Gln Glu Gln Thr Gly Asn Lys Arg Tyr Thr Phe Ile20 25 30Ala Ser Lys Leu Gln Glu Gln Thr Gly Asn Lys Arg Tyr Thr Phe Ile20 25 30

caa gca cat aga gct tgg ctt ctc aaa tta atg ttt cta tgg att ctt 144Gln Ala His Arg Ala Trp Leu Leu Lys Leu Met Phe Leu Trp Ile Leu35 40 45caa gca cat aga gct tgg ctt ctc aaa tta atg ttt cta tgg att ctt 144Gln Ala His Arg Ala Trp Leu Lys Leu Met Phe Leu Trp Ile Leu35 40 45

ctg atg gct ctg att agt cgt ate ate tac age aaa atg gat gtg ggt 192Leu Met Ala Leu Ile Ser Arg Ile Ile Tyr Ser Lys Met Asp vai Glyctg atg gct ctg att agt cgt up to tac age aaa atg gat gtg ggt 192Leu Met Wing Leu Ile Ser Arg Ile Ile Tyr Ser Lys Met Asp Gly Gly

50 55 6050 55 60

act aaa gtg aga agg aaa gag gtt ttg ggt agt ctt tgt gat caa agg 240Thr Lys Val Arg Arg Lys Glu Val Leu Gly Ser Leu Cys Asp Gln Arg65 70 75 80act aaa gtg aga aga aaa gag gtt ttg ggt agt ctt tgt gat caa agg 240Thr Lys Val Arg Arg Lys Glu Val Leu Gly Ser Leu Cys Asp Gln Arg65 70 75 80

gct aga atg ttg caa gac caa ttc agt gtt agt gtc aac cat gtt cat 288Ala Arg Met Leu Gln Asp Gln Phe Ser Val Ser vai Asn His vai Hisgct aga atg ttg caa gac caa ttc agt gtt agt gtc aac cat gtt cat 288Ala Arg Met Leu Gln Asp Gln Phe Ser Val Ser Go Asn His Go His

85 90 9585 90 95

gct ctt gcc ate ctt gtt tca act ttc cat tat tac aga aac cct tet 336Ala Leu Ala Ile Leu Val Ser Thr Phe His Tyr Tyr Arg Asn Pro Ser100 105 110gct ctt gcc until ctt gtt tca act ttc cat tat tac aga aac cct tet 336Ala Leu Ala Ile Leu Val Ser Thr Phe His Tyr Arg Asn Pro Ser100 105 110

gcc att gac cag gtttgtgctt gaaagttttg atcattctgt gttggaaatg 388gcc att gac cag gtttgtgctt gaaagttttg atcattctgt gttggaaatg 388

Ala Ile Asp Gln115Wing Ile Asp Gln115

aaaaatactc aatctttgtt gtgtttttga acctttatgt atcctctgat gattatttga 448tgatttccct tcag gaa act ttt gca gaa tat acg gct agg acc gct ttc 498Glu Thr Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe120 125aaaaatactc aatctttgtt gtgtttttga acctttatgt atcctctgat gattatttga 448tgatttccct tcag gaa act ttt gca gaa tat acg gct agg acc gct ttc 498Glu Thr Phe Ala Glu Tyr Thr Ala Arg Thr Phe20

gaa agg ccg cta ctt agt gga gtg gcc tat gca caa aga gtt gtt aac 546 Glu Arg Pro Leu Leu Ser Gly Val Ala Tyr Ala Gln Arg Val Val Asn130 135 140gaa agg ccg cta ctt agt gga gtg gcc tat gca caa aga gtt gtt aac 546 Glu Arg Pro Leu Read Ser Gly Val Wing Tyr Wing Gln Arg Val Val Asn130 135 140

tcg gaa aga gag cag ttt gag aag cag cat gga gtg gtt ata aag aca 594ser Glu Arg Glu Gln Phe Glu Lys Gln His Gly Val Val Ile Lys Thr145 150 155 160tcg gaa aga gag cag ttt gag aag cag cat gga gtg gtt ata aag aca 594ser Glu Arg Glu Gln Phe Glu Lys Gln His Gly Val Val Ile Lys Thr145 150 155 160

atg gaa aga gag gct tca ccg gtt agg gat gag tat gca ccg gtc ata 642Met Glu Arg Glu Ala Ser Pro vai Arg Asp Glu Tyr Ala Pro vai Ile165 170 175ttt gct cag gaa act gtc tct tac ctt gag tct att gat atg atg tct 690Phe Ala Gln Glu Thr Val Ser Tyr Leu Glu Ser Ile Asp Met Met Seratg gaa aga gag gct tca ccg gtt agg gat gag tat gca ccg gtc ata 642Met Glu Arg Glu Ala Ser Pro Go Arg Asp Glu Tyr Ala Pro Go Ile165 170 175ttt gct cag gaa act gtc tac tct att gat atg tct 690P Gln Wing Glu Thr Val Ser Tyr Leu Glu Ser Ile Asp Met Met Ser

180 185 190180 185 190

gga gag gtaaagaacg acacttgtga accattccac cgcttgtttt tttttttttt 746gga gag gtaaagaacg acacttgtga accattccac cgcttgtttt tttttttttt 746

Gly GluGly Glu

tttttggtgg tggattactt ttattactac aatttctcat ttttgcaatg cag gag 802tttttggtgg tggattactt ttattactac aatttctcat ttttgcaatg cag gag 802

Glu195Glu195

gat cga gag aac ata atg aga gct aga gct act ggg aaa gct gtt ctg 850Asp Arg Glu Asn Ile Met Arg Ala Arg Ala Thr Gly Lys Ala Val Leugat cga gag aac ata atg aga gct aga gct act ggg aaa gct gtt ctg 850Asp Arg Glu Asn Ile Met Arg Wing Arg Wing Thr Gly Lys Wing Val Leu

200 205 210200 205 210

act age cct ttt agg ttg ttg ggt tct cat cat ctc ggt gtg gtt tta 898Thr Ser Pro Phe Arg Leu Leu Gly Ser His His Leu Gly Val Val Leu 215 220 225act age cct ttgg ttg ttg ggt tct cat cat ctc ggt gtg gtt tta 898Thr Ser Pro Phe Arg Leu Leu Gly Ser His His Leu Gly Val Val Leu 215 220 225

aca ttt cct gtt tac aaa tct aag ctc cct ccc aac cca aca aca gaa 946Thr Phe Pro vai Tyr Lys Ser Lys Leu Pro Pro Asn Pro Thr Thr Gluaca ttt cct gt tac aaa tct aag ctc ccc aac cca aca aca gaa 946Thr Phe Pro will Tyr Lys Ser Lys Leu Pro Pro Asn Pro Thr Thr Glu

230 235 240230 235 240

gag ctc att aaa gcg acc gea gg gtatatgctt atttcactaa ttgtcagctt 999Glu Leu Ile Lys Ala Thr Ala Gly245 250gag ctc att aaa gcg acc gea gg gtatatgctt atttcactaa ttgtcagctt 999Glu Leu Ile Lys Ala Thr Ala Gly245 250

tttgttttta gaatcttttt tcttatgttt catattagta attagaggat gaaggttgtg 1059tttagtataa aaattttagg tacattatta gcattttatt aagcagcagt taatctgtca 1119atgccactat tagtgttatc gacacaactt ctatcattga gttgaagaaa actgtttctg 1179ttacaaggct tgtatttggt tatattttgt tgtaagcatt tcaatagaac atgtgattaa 1239tgaacttttt aattggaggc ag a tat gtt gga gga tcc ttt gat gtg gag 1289tttgttttta gaatcttttt tcttatgttt catattagta attagaggat gaaggttgtg 1059tttagtataa aaattttagg tacattatta gcattttatt aagcagcagt taatctgtca 1119atgccactat tagtgttatc gacacaactt ctatcattga gttgaagaaa actgtttctg 1179ttacaaggct tgtatttggt tatattttgt tgtaagcatt tcaatagaac atgtgattaa 1239tgaacttttt aattggaggc g tat gga gga tcc gtt gtg gag gat ttt 1289

Tyr vai Gly Gly Ser Phe Asp Val Glu255 260Tyr will Gly Gly Be Phe Asp Val Glu255 260

tca ctt gtg gaa aat tta ttt ggt caa ctt gct ggt cat caa gea att 1337ser Leu vai Glu Asn Leu Phe Gly Gln Leu Ala Gly His Gln Ala Iletca ctt gtg gaa aat tta ttt ggt caa ctt gct ggt cat caa gea att 1337ser Leu goes Glu Asn Leu Phe Gly Gln Leu Ala Gly His Gln Ala Ile

265 270 275265 270 275

ttg gtc aat gta tat gat gtc acg aac tct tct gat ccc cta ate atg 1385Leu Val Asn Val Tyr Asp Val Thr Asn Ser Ser Asp Pro Leu Ile Metttg gtc aat gta tat gat gtc acg aac tct tct gat ccc cta till atg 1385Leu Val Asn Val Tyr Asp Val Thr Asn Ser Ser Asp Pro Leu Ile Met

280 285 290280 285 290

tat ggc aac caa tac gaa gaa ggt gat gtt tct ctt gtc cat gaa agt 14B3Tyr Gly Asn Gln Tyr Glu Glu Gly Asp Val Ser Leu Val His Glu Ser 295 300 305tat ggc aac caa tac gaa ga ggt gat gtt tct ctt gtc cat gaa agt 14B3Tyr Gly Asn Gln Tyr Glu Glu Gly Asp Val Ser Leu Val His Glu Ser 295 300 305

aag ctt gat ttt gga gat cca tac agg aaa cat caa atg ata tgt ag 1480Lys Leu Asp Phe Gly Asp Pro Tyr Arg Lys His Gln Met Ile Cys Argaag ctt gat ttt gga gat cca tac agg aaa cat caa atg ata tgt ag 1480Lys Leu Asp Phe Gly Asp Pro Tyr Arg Lys His Gln Met Ile Cys Arg

310 315 320310 315 320

gtaggttctt ctggctactg tacattacat tttctggaac tgactttcgg cattttgtca 1540gcagaaagat gcttatgatc aaacttttgt acag g tat cac cag aag gca ccg 1593gtaggttctt ctggctactg tacattacat tttctggaac tgactttcgg cattttgtca 1540gcagaaagat gcttatgatc aaacttttgt acag g tat cac cag aag gca ccg 1593

Tyr His Gln Lys Ala Pro325 330Tyr His Gln Lys Wing Pro325 330

cca aat tgg aca gca ctt tct act gca ate cta ttc ttt gtg att ctt 1641Pro Asn Trp Thr Ala Leu Ser Thr Ala Ile Leu Phe Phe vai Ile Leu 335 340 345cca aat tgg aca gca ctt tct act gca until cta ttc ttt gtg att ctt 1641Pro Asn Trp Thr Ala Leu Ser Thr Ala Ile Leu Phe Phe Go Ile Leu 335 340 345

ctt tta ate ggt tac att tta tat ggt gct ggg aat cat att gtc aaa 1689Leu Leu Ile Gly Tyr Ile Leu Tyr Gly Ala Gly Asn His Ile vai Lysctt tta till ggt tac att tta tat ggt gct ggg aat cat att gtc aaa 1689Leu Leu Ile Gly Tyr Ile Leu Tyr Gly Ala Gly Asn His Ile Goes Lys

350 355 360350 355 360

gta gag gat gat ttc cac gaa atg cag gag cta aaa gtt cga gca gag 1737Val Glu Asp Asp Phe His Glu Met Gln Glu Leu Lys vai Arg Ala Glu365 370 375gta gag gat gat ttc cac gaa atg cag gag cta aaa gtt cga gca gag 1737Val Glu Asp Asp Phe His Glu Met Gln Glu Leu Lys Goes Arg Wing Glu365 370 375

gca gcc gat gtt gcc aag tca cag gtacttttcc ttgacatgtc gttggcactt 1791Ala Ala Asp Val Ala Lys Ser Gln380 385gca gcc gat gtt gcc aag tca cag gtacttttcc ttgacatgtc gttggcactt 1791Ala Wing Asp Val Wing Lys Ser Gln380 385

ggcatatttc gatatttcct ttgattacgc aacaataact aatactgaaa tttttgtttt 1851tgtcag ttt cta gct act gtc tct cat gaa att aga aca cct atg aat 1899Phe Leu Ala Thr Val Ser His Glu Ile Arg Thr Pro Met Asn390 395 400ggcatatttc gatatttcct ttgattacgc aacaataact aatactgaaa tttttgtttt 1851tgtcag ttt cta gct act gtc cat gaa att aga aca cct atg aat 1899Phe Leu Ala Thr Val Be His Glu Ile Arg Thr Pro Met Asn390 395 400

ggc att tta g gtaactgcaa aattctctct ctctttgcct aacatgtaac 1949Gly Ile Leuggc att tta g gtaactgcaa aattctctct ctctttgcct aacatgtaac 1949Gly Ile Leu

atatgctttc atagctttca cttgctcatt aataatgtta tcacaaaaca aaactctgta 2009tgattgtag ga atg ctt ggt ctg ctt cta cgc acg gaa ttg aac tca act 2059Gly Met Leu Gly Leu Leu Leu Arg Thr Glu Leu Asn Ser Thr405 410 415atatgctttc atagctttca cttgctcatt aataatgtta tcacaaaaca aaactctgta 2009tgattgtag ga atg ctt ggt ctg ctt cta cgc acg gaa ttg aac tca act 2059Gly Met Leu Gly Leu Leu Leu Arg Thr Thr515

caa cgg gac tat gct cag act gct caa gca tgt ggg aag gct ctg ata 2107Gln Arg Asp Tyr Ala Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu Ile420 425 430caa cgg gac tat gct cag act gct caa gca tgt ggg aag gct ctg ata 2107Gln Arg Asp Tyr Ala Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu Ile420 425 430

gca tta ata aat gaa gtg ctt gac cga gca aaa att gaa gct ggc aaa 2155Ala Leu Ile Asn Glu vai Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys435 440 445gca tta ata aat gaa gtg ctt gac cga gca aaa att gaa gct ggc aaa 2155Ala Leu Ile Asn Glu Goes Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys435 440 445

tta gag ctg gaa gcg gtt cca ttt gac ctt cgt tcc ata ctc gat gat 2203Leu Glu Leu Glu Ala Val Pro Phe Asp Leu Arg Ser Ile Leu Asp Asp450 455 460 465tta gag ctg gaa gcg gtt cca ttt gac ctt cgt tcc ata ctc gat gat 2203Leu Glu Leu Glu Ala Val Pro Phe Asp Leu Arg Ser Ile Leu Asp Asp450 455 460 465

gtc ctc tct ctt ttt tca gag aag tct aga cac aaa ggt tta gag 2248gtc ctc tct ttt tca gag aag tct aga cac aaa ggt tta gag 2248

Val Leu Ser Leu Phe Ser Glu Lys Ser Arg His Lys Gly Leu Glu 470 475 480Val Leu Be Leu Phe Be Glu Lys Be Arg His Lys Gly Leu Glu 470 475 480

gtatgctcaa tcgttactaa acctattgtg aaggccaata cgagtttcat aataatatct 2308taattcggtt tcctctcttt atttgatgat ctaacttcat tatccttttt ttttttttca 2368ttttaccttt catag ctg gca gtg ttt gtt tct gat aaa gtt cca gat att 2419Leu Ala Val Phe vai Ser Asp Lys Val Pro Asp Ile485 490gtatgctcaa tcgttactaa acctattgtg aaggccaata cgagtttcat aataatatct 2308taattcggtt tcctctcttt atttgatgat ctaacttcat tatccttttt ttttttttca 2368ttttaccttt catag ctg gca gtg ttt GTT GTT TCT aaa gat gat cca att 2419Leu Ala Val Phe will be Asp Lys Asp Val Pro Ile485 490

gtt atg gga gat cct ggg aga ttc aga caa att gta aca aat ctt gtt 2467vai Met Gly Asp Pro Gly Arg Phe Arg Gln Ile Val Thr Asn Leu Valgtt atg gga gat cct ggg aga ttc aga caa att gta aca aat ctt gtt 2467vai Met Gly Asp Pro Gly Arg Phe Arg Gln Ile Val Thr Asn Leu Val

495 500 505495 500 505

ggc aac tct gtt aaa gtaagtggaa ttttcaaatt ttatttgcct aaagttattt 2522Gly Asn Ser Val Lys510ggc aac tct gtt aaa gtaagtggaa ttttcaaatt ttatttgcct aaagttattt 2522Gly Asn Ser Val Lys510

gcattactaa tattcatctt atgttggaaa tgtttgaaat tttgctgttg tatagcaaga 2582tacttgttga ttacatttct gattaacttc aatatattgc ag ttc act gag cga 2636gcattactaa tattcatctt atgttggaaa tgtttgaaat tttgctgttg tatagcaaga 2582tacttgttga ttacatttct gattaacttc aatatattgc ag ttc act gag cga 2636

Phe Thr Glu ArgPhe Thr Glu Arg

515515

ggc cat ata ttt gtt aaa gtt cat tta tct gaa aac aga aag ccc gta 2684Gly His lie Phe vai Lys Val His Leu Ser Glu Asn Arg Lys Pro vai520 525 530ggc cat ata ttt gtt aaa gtt cat tta tct gaa aac aga aag ccc gta 2684Gly his lie phe will lys val his leu be glu asn arg lys pro vai520 525 530

aca aat gga aag cat gag act tat cga aat gga ggg tct gaa gaa gtt 2732Thr Asn Gly Lys His Glu Thr Tyr Arg Asn Gly Gly Ser Glu Glu vai535 540 545aca aat gga aag cat gag act tat cga aat gga ggg tct gaa gaa gtt 2732Thr Asn Gly Lys His Glu Thr Tyr Arg Asn Gly Gly Ser Glu Glu vai535 540 545

gtg cat gca tct ggc ggt tat aat ctc aaa acg cta agt gga tat gaa 2780vai His Ala Ser Gly Gly Tyr Asn Leu Lys Thr Leu Ser Gly Tyr Glu550 555 560 565gtg cat gca tct ggc ggt tat aat ctc aaa acg cta agt gga tat gaa 2780vai His Ala Ser Gly Gly Tyr Asn Leu Lys Thr Leu Ser Gly Tyr Glu550 555 560 565

gct gct gat gaa cgc aac aac tgg gat aat ttt aac cac tta att gct 2828Ala Ala Asp Glu Arg Asn Asn Trp Asp Asn Phe Asn His Leu Ile Ala570 575 580gct gct gat gaa cgc aac aac tgg gat aat ttt aac cac tta att gct 2828Ala Ala Asp Glu Arg Asn Asp Trp Asp Asn Phe Asn His Leu Ile Ala570 575 580

gat gaa gag ttt ttc tgc gat gct tca act aaa aaa gtg gcc tcg aat 2876Asp Glu Glu Phe Phe Cys Asp Ala Ser Thr Lys Lys vai Ala Ser Asngat gaa gag ttt ttc tgc gat gct tca act aaa aaa gtg gcc tcg aat 2876Asp Glu Glu Phe Phe Cys Asp Ala Ser Thr Thr Lys Lys Go Ala Ser Asn

585 590 595585 590 595

gaa ttt tat gaa caa gtc acc ttg atg gtc tgt gtc gaa gac act gga 2924Glu Phe Tyr Glu Gln Val Thr Leu Met Val Cys vai Glu Asp Thr Gly600 605 610gaa ttt tat gaa caa gtc acc ttg atg gtc tgt gtc gaa gac act gga 2924Glu Phe Tyr Glu Gln Val Thr Leu Met Val Cys Goes Glu Asp Thr Gly600 605 610

att gga att cct ttc tcg gcc caa gat agg att ttc atg cct ttt gtt 2972Ile Gly Ile Pro Phe Ser Ala Gln Asp Arg Ile Phe Met Pro Phe Val615 620 625att gga att cct ttc tcg gcc caa gat agg att ttc atg cct ttt gtt 2972Ile Gly Ile Pro Phe Ser Ala Gln Asp Arg Ile Phe Met Pro Phe Val615 620 625

cag gca gat age tcg act tct aga aat tat ggt ggt acc ggc att ggt 3020Gln Ala Asp Ser Ser Thr Ser Arg Asn Tyr Gly Gly Thr Gly Ile Gly630 635 640 645cag gca gat age tcg act tct aga aat tat ggt ggt acc ggc att ggt 3020Gln Wing Asp Be Ser Thr Be Arg Asn Tyr Gly Gly Thr Gly Ile Gly630 635 640 645

ttg agt ate agt aag tgc cta gtt gaa cta atg ggt ggt caa ata aac 3068Leu Ser Ile Ser Lys Cys Leu vai Glu Leu Met Gly Gly Gln Ile Asn650 655 660ttg agt till agt aag tgc cta gtt gaa cta atg ggt ggt caa ata aac 3068Leu Ser Ile Ser Lys Cys Leu goes Glu Leu Met Gly Gly Gln Ile Asn650 655 660

ttt ata age cgg ccg cag gtt gga age acc ttt tca ttt act gcg gat 3116Phe Ile ser Arg Pro Gln Val Gly Ser Thr Phe Ser Phe Thr Ala Aspttt ata age cgg ccg cag gtt gga age acc ttt tca ttt act gcg gat 3116Phe Ile Be Arg Pro Gln Val Gly Be Thr Phe Be Phe Thr Wing Asp

665 670 675665 670 675

ttt gga ata ttt aag aag aat cca ata act gag gtg aag aag gtt aac 3164Phe Gly Ile Phe Lys Lys Asn Pro Ile Thr Glu vai Lys Lys Val Asn680 685 690ttt gga ata ttt aag aag aat cca ata act gag gtg aag aag gtt aac 3164Phe Gly Ile Phe Lys Lys Asn Pro Ile Thr Glu Goes Lys Lys Val Asn680 685 690

tat gaa gat cta cca tcc agt ttt aga ggg ctt aaa gcc gtt gtg gtt 3212Tyr Glu Asp Leu Pro Ser Ser Phe Arg Gly Leu Lys Ala Val Val Valtat gaa gat cta cca tcc agt ttt aga ggg ctt aaa gcc gtt gtg gtt 3212Tyr Glu Asp Leu Pro To Be Phe Arg Gly Leu Lys Ala Val Val Val Val

695 700 705695 700 705

gat ggg aaa cct gtt aga gct gct gtg act aga tac cat ttg aag aga 3260Asp Gly Lys Pro Val Arg Ala Ala vai Thr Arg Tyr His Leu Lys Arg 710 715 720 725gat ggg aaa cct gtt aga gct gct gtg act aga tac cat ttg aag aga 3260Asp Gly Lys Pro Val Arg Wing Wing Go Thr Arg Tyr His Leu Lys Arg 710 715 720 725

ctt ggg ata caa gtt aaa gtc gca aat gcc ate aat aag gct gtt tcc 3308Leu Gly Ile Gln Val Lys Val Ala Asn Ala Ile Asn Lys Ala Val Serctt ggg ata caa gtt aaa gtc gca aat gcc until aat aag gct gtt tcc 3308Leu Gly Ile Gln Val Lys Val Ala Asn Ala Ile Asn Lys Ala Val Ser

730 735 740730 735 740

ttg tgt ggg aaa aat ggg gct tcc age aca gg gtaagttttt aattttcctt 3360 Leu Cys Gly Lys Asn Gly Ala ser Ser Thr Gly745 750ttg tgt ggg aaa aat ggg gct tcc age aca gg gtaagttttt aattttcctt 3360 Leu Cys Gly Lys Asn Gly Ala Ser Ser Thr Gly745 750

tttgtaatta atgcattgct atttttaatg aattaatgtt aggactatgt gtgtttaatc 3420ataattccat ctaaagtcac acatgatagg ctgtgacttg ctgacagtgt aaaacgtata 3480aaacattata cactaacagt gtatcaaaat taaactcttt gaagaattgc atttgttctt 3540 gttgtgtggt tcaatacttg agttcattaa ctgagttaaa atatttggtt gtggcag g 3598tta ttc cag cct gat att att ttt gtt gag aaa gat tca tgg gtt tgt 3646Leu Phe Gln Pro Asp Ile Ile Phe Val Glu Lys Asp Ser Trp vai Cystttgtaatta atgcattgct atttttaatg aattaatgtt aggactatgt gtgtttaatc 3420ataattccat ctaaagtcac acatgatagg ctgtgacttg ctgacagtgt aaaacgtata 3480aaacattata cactaacagt gtatcaaaat taaactcttt gaagaattgc atttgttctt 3540 gttgtgtggt tcaatacttg agttcattaa ctgagttaaa atatttggtt gtggcag g 3598tta ttc cag cct gat att att ttt gtt gag aaa gat tca tgg gtt tgt 3646Leu Phe Gln Pro Asp Ile Ile Phe Val Glu Lys Asp Ser Trp Goes Cys

755 760 765755 760 765

gga gag gac ggg ate ttc agt gtg cgc caa ctg gac tgg aaa cag aat 3694 Gly Glu Asp Gly Ile Phe Ser vai Arg Gln Leu Asp Trp Lys Gln Asn770 775 780gga gag gac ggg till ttc agt gtg cgc caa ctg gac tgg aaa cag aat 3694 Gly Glu Asp Gly Ile Phe Ser goes Arg Gln Leu Asp Trp Lys Gln Asn770 775 780

gga cat ata ttt aag atg cct caa atg ate ctt cta gct aca aat att 3742Gly His lie Phe Lys Met Pro Gln Met Ile Leu Leu Ala Thr Asn Ile785 790 795 800gga cat ata ttt aag atg cct hunt atg until ctt cta gct aca aat att 3742Gly His Lie Phe Lys Met Pro Gln Met Ile Leu Leu Thr Asn Ile785 790 795 800

agt aat gac gaa ttt gat aaa gct aaa tcc gca ggt ttt agt gat acg 3790Ser Asn Asp Glu Phe Asp Lys Ala Lys Ser Ala Gly Phe Ser^Asp Thragt aat gac gaa ttt gat aaa gct aaa tcc gca ggt ttt agt gat acg 3790Ser Asn Asp Glu Phe Asp Lys Ala Lys Ser Ala Gly Phe Ser ^ Asp Thr

805 810 815805 810 815

gtg ate atg aag cca ctg aga gct agt atg gtg gga gct tgc ctt cag 3838Val lie Met Lys Pro Leu Arg Ala Ser Met Val Gly Ala Cys Leu Gln 820 825 830gtg till atg aag cca ctg aga gct agt atg gtg gga gct tgc ctt cag 3838Val lie Met Lys Pro Leu Arg Wing Ser Met Val Gly Ala Cys Leu Gln 820 825 830

caa gtt ttg gga aca ggc aag aag aga cag ctg gga aaa gag atg cct 3886Gln vai Leu Gly Thr Gly Lys Lys Arg Gln Leu Gly Lys Glu Met Pro835 840 845caa gtt ttg gga aca ggc aag aag aga cag ctg gga aaa gag atg cct 3886Gln will Read Gly Thr Gly Lys Lys Arg Gln Read Gly Lys Glu Met Pro835 840 845

aat ggt tca act tca gtt cga age ctt ctg ttc gga aag aaa att tta 3934ag ggt tca act tca gtt cga age ctt ctg ttc gga aag aaa att tta 3934

Asn Gly Ser Thr Ser vai Arg Ser Leu Leu Phe Gly Lys Lys Ile LeuAsn Gly Be Thr Be Go Arg Be Read Leu Phe Gly Lys Lys Ile Leu

850 855 860850 855 860

gtg gtt gat gac aat gta gta aac cgg agg gtg gct gea ggt gcc ttg 3982gt gtt gat gac aat gta gta aac cgg agg gtg gct gea ggt gcc ttg 3982

vai vai Asp Asp Asn Val Val Asn Arg Arg vai Ala Ala Gly Ala Leugo go Asp Asp Asn Val Val Asn Arg Arg go Ala Wing Gly Ala Leu

865 870 875 880865 870 875 880

aaa aac ttt gga gcg gat gtg aag tgt gea gat age ggc aaa gct gct 4030aaa aac ttt gga gcg gat gtg aag tgt gea gat age ggc aaa gct gct 4030

Lys Asn Phe Gly Ala Asp Val Lys Cys Ala Asp Ser Gly Lys Ala AlaLys Asn Phe Gly Wing Asp Val Lys Cys Wing Asp Ser Gly Lys Wing

885 890 895885 890 895

ctt gaa atg ctt caa ttc cct cac aag ttt gat gct tgc ttc atg gat 4078ct gaa atg ct caa ttc cct cac aag ttt gat gct tgc ttc atg gat 4078

Leu Glu Met Leu Gln Phe Pro His Lys Phe Asp Ala Cys Phe Met AspRead Glu Met Read Le Gln Phe Pro His Lys Phe Asp

900 905 910900 905 910

att caa atg cca gaa atg gac gg gtatgtttgt ttgcatcgat tattaatttt 4131att caa atg cca gaa atg gac gg gtatgtttgt ttgcatcgat tattaatttt 4131

Ile Gln Met Pro Glu Met Asp GlyGlle Met Pro Glu Met Asp Gly

915915

ttttttggta ttatttgaac ataacatgtt aacatgatag ttgcattgct gataataaaa 4191aaaaaaactt gcataagttg aatgcaagtt acgtgatagt tgccctgctg ataaattcaa 4251tgatatgcac gtaagttgaa tgcacgtaac ttgcatgttg gctatactaa ttcacattca 4311aattattttg tttgtatata aactctaatt acacatgtta tgtatgttct aattcacatc 4371catactatat ttgtttgtat ataaacctta tcaatagatg ttatgtgtgt tctaatccac 4431atgaatatta tttttacgca tattgttttt tataaatcat gtacttgcat tattttcag 4490g ttt gaa gea act cgt cga att cgg gag atg gag agg aca gcg aat gag 4539ttttttggta ttatttgaac ataacatgtt aacatgatag ttgcattgct gataataaaa 4191aaaaaaactt gcataagttg aatgcaagtt acgtgatagt tgccctgctg ataaattcaa 4251tgatatgcac gtaagttgaa tgcacgtaac ttgcatgttg gctatactaa ttcacattca 4311aattattttg tttgtatata aactctaatt acacatgtta tgtatgttct aattcacatc 4371catactatat ttgtttgtat ataaacctta tcaatagatg ttatgtgtgt tctaatccac 4431atgaatatta tttttacgca tattgttttt tataaatcat gtacttgcat tattttcag 4490g ttt gaa g and act cgt cga att cgg gag atg gag agg aca gcg aat gag 4539

Phe Glu Ala Thr Arg Arg Ile Arg Glu Met Glu Arg Thr Ala Asn GluPhe Glu Wing Thr Arg Arg Ile Arg Glu Met Glu Arg Thr Wing Asn Glu

925 930 935925 930 935

gag acg aat agt gaa tgt ggt gaa agg aaa agt gaa ttc cat tta cct 4587gag acg aat agt gaa tgt ggt gaa agg aaa agt gaa ttc cat tta cct 4587

Glu Thr Asn Ser Glu Cys Gly Glu Arg Lys Ser Glu Phe His Leu ProGlu Thr Asn Be Glu Cys Gly Glu Arg Lys Be Glu Phe His Leu Pro

940 945 950940 945 950

ata ttg gcc atg aca gea gat gta ate cat gct aca tat gaa gag tgt 4635ata ttg gcc atg aca gea gat gta until cat gct aca tat gaa gag tgt 4635

Ile Leu Ala Met Thr Ala Asp Val Ile His Ala Thr Tyr Glu Glu CysIle Leu Wing Met Thr Wing Asp Val Ile His Wing Thr Tyr Glu Glu Cys

955 960 965955 960 965

ttg aaa tgt ggg atg gat ggt tat gtt tca aaa cct ttt gag gaa gag 4683Leu Lys Cys Gly Met Asp Gly Tyr vaittg aaa tgt ggg atg gat ggt tat gtt tca aaa cct ttt gag gaa gag 4683Leu Lys Cys Gly Met Asp Gly Tyr will

970 975970 975

aat ctt tat caa gca gtt gca aag tttAsn Leu Tyr Gln Ala vai Ala Lys Phe985 990aat ctt tat caa gca gtt gca aag tttAsn Read Tyr Gln Wing Go Wing Lys Phe985 990

gta gat tca tgaVal Asp sergta gat tca tgaVal Asp be

Ser Lys Pro Phe Glu Glu Glu980Ser Lys Pro Phe Glu Glu Glu980

ttc cag aca aaa cct act tca 4731Phe Gln Thr Lys Pro Thr Ser995 1000ttc cag aca aaa cct act tca 4731Phe Gln Thr Lys Pro Thr Ser995 1000

47434743

<210> 8 <211> 3694 <212> DNA <213> Arabidopsis thal <220> <221> 5'UTR <222> (1)..(68) <220> <221> CDS <222> (69)..(3311) <220> <221> 3'UTR <222> (3312)..(3694) <220> <221> misc_feature <222> (3594)..(3594) <223> η is a, c, g, or <220> <221> misc_feature <222> (3631)..(3631) <223> η is a, c, g, or <400> 8<210> 8 <211> 3694 <212> DNA <213> Arabidopsis thal <220> <221> 5'UTR <222> (1) .. (68) <220> <221> CDS <222> (69) .. (3311) <220> <221> 3'UTR <222> (3312) .. (3694) <220> <221> misc_feature <222> (3594) .. (3594) <223> η is a, c, g, or <220> <221> misc_feature <222> (3631) .. (3631) <223> η is a, c, g, or <400> 8

aaaaaatctc actaaaacaa aagaagaaga aagaagaaag aaaatggaat acctacattt 60ttgaagtg atg aga aga gat ttt gtg tat aat aat aat gca atg ttc aat 11010aaaaaatctc actaaaacaa aagaagaaga aagaagaaag aaaatggaat acctacattt 60ttgaagtg atg aga aga gat ttt gtg tat aat aat gca atg ttc aat 11010

1515

cctcct

ProPro

1515

catcat

HisHis

gaggag

GluGlu

ggtGlyggtGly

cgtArgcgtArg

cagcag

GlnGln

9595

atgatg

MetMet

tta25 Leutta25 Leu

agtSeragtSer

2020

3030

gagGlugagGlu

Met Arg Arg Asp Phe vai Tyr Asn Asn Asn Ala Met Phe Asn15 10Met Arg Arg Asp Phe Go Tyr Asn Asn Asn Wing Met Phe Asn15 10

ctc aca act cat tac age tca gat atg aac tgg gea ctc aac aat 158Leu Thr Thr His Tyr ser ser Asp Met Asn Trp Ala Leu Asn Asn20 25 30ctc aca act cat tac act tca gat atg aac tgg gea ctc aac aat 158Leu Thr Thr His Tyr be ser Asp Met Asn Trp Ala Leu Asn Asn20 25 30

caa gaa gaa gaa gaa gag cca cga aga att gaa att tet gat tcc 206Gln Glu Glu Glu Glu Glu Pro Arg Arg Ile Glu Ile Ser Asp Sercaa gaa gaa gaa ga ga cca cga cga aga att gaa att tet gat tcc 206Gln Glu Glu Glu Glu Glu Pro Arg Arg Ile Glu Ile Ser Asp Ser

35 40 4535 40 45

tca cta gaa aac ttg aaa age age gat ttt tat caa ctg ggt ggt 254Ser Leu Glu Asn Leu Lys Ser ser Asp Phe Tyr Gln Leu Gly Glytca cta gaa aac ttg aaa age age gat ttt tat caa ctg ggt ggt 254Ser Leu Glu Asn Leu Lys Being Asp Phe Tyr Gln Leu Gly Gly

50 55 6050 55 60

ggt gct ctg aat tcg tca gaa aag ccg aga aag ate gat ttt tgg 302Gly Ala Leu Asn Ser Ser Glu Lys Pro Arg Lys Ile Asp Phe Trpggt gct ctg aat tcg tca gaa aag ccg aga aag till gat ttt tgg 302Gly Ala Leu Asn Ser Ser Glu Lys Pro Arg Lys Ile Asp Phe Trp

65 70 7565 70 75

tcg ggg ttg atg ggt ttt gcg aag atg cag cag cag caa cag ctt 350Ser Gly Leu Met Gly Phe Ala Lys Met Gln Gln Gln Gln Gln Leu80 85 90tcg ggg ttg atg ggt ttt gcg aag atg cag cag cag caa cag ctt 350Ser Gly Leu Met Gly Phe Ala Lys Met Gln Gln Gln Gln Leu80 85 90

cat tca gtg gcg gtg aag atg aac aat aat aat aat aac gat cta 398His Ser Val Ala vai Lys Met Asn Asn Asn Asn Asn Asn Asp Leu100 105 110cat tca gtg gcg gtg aag atg aac aat aat aat aat aat gat cta 398His Ser Val Wing Go Lys Met Asn Asn Asn Asn Asn Asn Asn Asn Leu100 105 110

ggt aat aaa aaa ggg tca act ttc ata caa gaa cat cga gea ttg 446Gly Asn Lys Lys Gly Ser Thr Phe Ile Gln Glu His Arg Ala Leuggt aat aaa aaa ggg tca act ttc ata caa gaa cat cga gea ttg 446Gly Asn Lys Lys Gly Ser Thr

115 120 125115 120 125

cca aaa gct ttg att ctg tgg ate ate att gtt ggg ttt ata age 494Pro Lys Ala Leu Ile Leu Trp Ile Ile Ile Val Gly Phe Ile Sercca aaa gct ttg att ctg tgg until until att gtt ggg ttt ata age 494Pro Lys Ala Leu Ile Leu Trp Ile Ile Ile Val Gly Phe Ile Ser

130 135 140130 135 140

ggg att tat cag tgg atg gat gat gct aat aag att aga agg gaa 542Gly Ile Tyr Gln Trp Met Asp Asp Ala Asn Lys Ile Arg Arg Gluggg att tat cg tgg atg gat gat gct aat aag att aga agg gaa 542Gly Ile Tyr Gln Trp Met Asp Asp Ala Asn Lys Ile Arg Arg Glu

145 150 155145 150 155

gtt ttg gtc age atg tgt gat caa aga gct aga atg ttg cag gat 590vai Leu vai Ser Met Cys Asp Gln Arg Ala Arg Met Leu Gln Asp160 165 170caa ttt agt gtt agt gtt aat cat gtt cat gct ttg gct att ctc gtc 638Gln Phe Ser Val Ser vai Asn His vai His Ala Leu Ala Ile Leu Val175 180 185 190gtt ttg gtc age atg tgt gat caa aga gct aga atg ttg cag gat 590vai Read will be Met Cys Asp Gln Arg Ala Arg Met Leu Gln Asp160 165 170caa ttt agt gtt agt gtt aat cat gtt ttg gct att ctc gtc P3838 Val Ser goes Asn His goes His Wing Leu Wing Ile Leu Val175 180 185 190

tcc act ttt cat tac cac aag aac cct tct gca att gat cag gag aca 686ser Thr Phe His Tyr His Lys Asn Pro Ser Ala Ile Asp Gln Glu Thr195 200 205tcc act ttt cat tac aag aac cct tct gca att gat cag gag aca 686ser Thr Phe His Tyr Lys Asn Pro Ser Ala Ile Asp Gln Glu Thr195 200 205

ttt gcg gag tac acg gca aga aca gca ttt gag aga ccg ttg cta agt 734Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe Glu Arg Pro Leu Leu Ser210 215 220ttt gcg gag tac acg gca aga aca gca ttt gag aga ccg ttg cta agt 734Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe Glu Arg Pro Leu Leu Ser210 215 220

gga gtg gct tat gct gaa aaa gtt gtg aat ttt gag agg gag atg ttt 782Gly vai Ala Tyr Ala Glu Lys Val Val Asn Phe Glu Arg Glu Met Phe225 230 235gga gtg gct tat gct gaa aaa gtt gtg aat ttt gag agg gag atg ttt 782Gly goes Ala Tyr Ala Glu Lys Val Val Asn Phe Glu Arg Glu Met Phe225 230 235

gag cgg cag cac aat tgg gtt ata aag aca atg gat aga gga gag cct 830Glu Arg Gln His Asn Trp vai Ile Lys Thr Met Asp Arg Gly Glu Pro240 245 250gag cgg cag cac aat tgg gtt ata aag aca atg gat aga gga gag cct 830Glu Arg Gln His Asn Trp will Ile Lys Thr Met Asp Arg Gly Glu Pro240 245 250

tca ccg gtt agg gat gag tat gct cct gtt ata ttc tct caa gat agt 878Ser Pro Val Arg Asp Glu Tyr Ala Pro Val Ile Phe Ser Gln Asp Ser255 260 265 270tca ccg gtt agg gat gag tat gct cct gtt ata ttc tct caa gat agt 878Ser Pro Val Arg Asp Glu Tyr Pro Val Ile Phe Ser Gln Asp Ser255 260 265 270

gtc tct tac ctt gag tca ctc gat atg atg tca ggc gag gag gat cgt 926Val Ser Tyr Leu Glu Ser Leu Asp Met Met Ser Gly Glu Glu Asp Arg275 280 285gtc tct tac ct gag tca ctc gat atg tg ggc gag gag gat cgt 926Val Ser Tyr Leu Glu Ser Leu Asp Met Met Ser Gly Glu Glu Asp Arg275 280 285

gag aat att ttg cga gct aga gaa acc gga aaa gct gtc ttg act age 974Glu Asn Ile Leu Arg Ala Arg Glu Thr Gly Lys Ala Val Leu Thr Ser290 295 300gag aat att ttg cga gct aga gaa acc gga aaa gct gtc ttg act 974Glu Asn Ile Leu Arg Wing Arg Glu Thr Gly Lys Wing Val Leu Thr Ser290 295 300

cct ttt agg ttg ttg gaa act cac cat ctc gga gtt gtg ttg aca ttc 1022Pro Phe Arg Leu Leu Glu Thr His His Leu Gly Val vai Leu Thr Phe305 310 315cct ttt agg ttg ttg gaa act cat cat ctc gga gtt gtg ttg aca ttc 1022Pro Phe Arg Leu Leu Glu Thr His Leu Gly Val Goes Leu Thr Phe305 310 315

cct gtc tac aag tct tct ctt cct gaa aat ccg act gtc gaa gag cgt 1070Pro vai Tyr Lys Ser Ser Leu Pro Glu Asn Pro Thr vai Glu Glu Arg320 325 330cct gtc tac aag tct tct ctt cct gaa aat ccg act gtc gaa gag cgt 1070Pro will Tyr Lys Ser Be Leu Pro Glu Asn Pro Thr will Glu Glu Arg320 325 330

att gca gcc act gca ggg tac ctt ggt ggt gcg ttt gat gtg gag tct 1118Ile Ala Ala Thr Ala Gly Tyr Leu Gly Gly Ala Phe Asp Val Glu Ser335 340 345 350att gca gcc act gca ggg tac ctt ggt ggt gcg ttt gat gtg gag tct 1118Ile Wing Wing Thr Wing Wing Gly Tyr Leu Gly Gly Wing Phe Asp Val Glu Ser335 340 345 345 350

cta gtc gag aat tta ttt ggt cag ctt gct ggt aac caa gca ata gtt 1166Leu vai Glu Asn Leu Phe Gly Gln Leu Ala Gly Asn Gln Ala Ile Val355 360 365cta gtc gag aat tta ttt ggt cag ctt gct ggt aac caa gca ata gtt 1166Leu goes Glu Asn Leu Phe Gly Gln Leu Ala Gly Asn Gln Ala Ile Val355 360 365

gtg cat gtg tat gat ate acc aat gca tca gat cca ctt gtc atg tat 1214Val His vai Tyr Asp Ile Thr Asn Ala Ser Asp Pro Leu Val Met Tyrgtg cat gtg tat gat up acc aat gca tca gat cca ctt gtc atg tat 1214Val His Go Tyr Asp Ile Thr Asn Ala Ser Asp Pro Leu Val Met Tyr

370 375 380370 375 380

ggt aat caa gat gaa gaa gcc gac aga tet ctc tet cat gag age aag 1262Gly Asn Gln Asp Glu Glu Ala Asp Arg Ser Leu ser His Glu ser Lys385 390 395ggt aat caa gat gaa gaa gcc gac aga tet ctc tet cat gag age aag 1262Gly Asn Gln Asp Glu Glu Wing Asp Arg Ser Leu Be His Glu Be Lys385 390 395

ctc gat ttt gga gac ccc ttc agg aaa cat aag atg ata tgc agg tac 1310Leu Asp Phe Gly Asp Pro Phe Arg Lys His Lys Met Ile Cys Arg Tyrctc gat ttt gga gac ccc ttc agg aaa cat aag atg ata tgc agg tac 1310Leu Asp Phe Gly Asp Pro Phe Arg Lys His Lys Met Ile Cys Arg Tyr

400 405 410400 405 410

cac caa aag gca cca ata cca ttg aat gtg ctc aca act gtg cca ttg 1358His Gln Lys Ala Pro Ile Pro Leu Asn Val Leu Thr Thr Val Pro Leu415 420 425 430cac caa aag gca cca ata cca ttg aat gtg ctc aca act gtg cca ttg 1358His Gln Lys Ala Pro Ile Pro Leu Asn Val Leu Thr Thr Val Pro Leu415 420 425 430

ttc ttt gcg att ggt ttc ttg gtg ggt tat ata ctg tat ggt gca gct 1406Phe Phe Ala Ile Gly Phe Leu Val Gly Tyr Ile Leu Tyr Gly Ala Ala435 440 445ttc ttt gcg att ggt ttc ttg gtg ggt tat ata ctg tat ggt gca gct 1406Phe Phe Ala Ile Gly Phe Leu Val Gly Tyr Ile Leu Tyr Gly Ala435 440 445

atg cac ata gta aaa gtc gaa gat gat ttc cat gaa atg caa gag ctt 1454Met His Ile vai Lys vai Glu Asp Asp Phe His Glu Met Gln Glu Leuatg cac ata gta aaa gtc gaa gat gat ttc cat gaa atg caa gag ctt 1454Met His Ile Will Lys Will Glu Asp Asp Phe His Glu Met Gln Glu Leu

450 455 460450 455 460

aaa gtg cga gca gaa gct gct gat gtc gct aaa tcg cag ttt ctt gct 1502Lys Val Arg Ala Glu Ala Ala Asp Val Ala Lys Ser Gln Phe Leu Ala465 470 475aaa gtg cga gca gaa gct gct gat gtc gct aaa tcg cag ttt ctt gct 1502Lys Val Arg Wing Glu Wing Wing Asp Val Wing Wing Lys Ser Gln Phe Leu Wing Wing465 470 475

acc gtg tet cac gag ate agg aca cca atg aat ggc att ctc gga atg 1550Thr Val ser His Glu Ile Arg Thr Pro Met Asn Gly lie Leu Gly Metacc gtg tet cac gag up agg aca cca atg aat ggc att ctc gga atg 1550Thr Val be His Glu Ile Arg Thr Pro Met Asn Gly lie Leu Gly Met

480 485 490480 485 490

ctt gct atg ctc cta gat aca gaa cta age tcg aca cag aga gat tac 1598Leu Ala Met Leu Leu Asp Thr Glu Leu Ser ser Thr Gln Arg Asp Tyr495 500 505 510ctt gct atg ctc cta gat aca gaa cta age tcg aca cag aga gat tac 1598Leu Ala Met Leu Leu Asp Thr Glu Leu Being Thr Gln Arg Asp Tyr495 500 505 510

gct caa acc gct caa gta tgt ggt aaa gct ttg att gca ttg ata aat 1646Ala Gln Thr Ala Gln Val Cys Gly Lys Ala Leu Ile Ala Leu Ile Asngct caa acc gct caa gta tgt ggt aaa gct ttg att gca ttg ata aat 1646Ala Gln Thr Wing Gln Val Cys Gly Lys Wing Leu Ile Wing Leu Ile Asn

515 520 525515 520 525

gag gtt ctt gat cgc gcc aag att gaa gct gga aag ctg gag ttg gaa 1694Glu Val Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys Leu Glu Leu Glu530 535 540gag gtt ctt gat cgc gcc aag att gaa gct gga aag ctg gag ttg gaa 1694Glu Val Leu Asp Arg Wing Lys Ile Glu Wing Gly Lys Leu Glu Leu Glu530 535 540

tca gta cca ttt gat ate cgt tca ata ttg gat gat gtc ctt tet cta 1742Ser Val Pro Phe Asp Ile Arg ser Ile Leu Asp Asp vai Leu Ser Leutca gta cca ttt gat till cgt tca ata ttg gat gat gtc ctt tet cta 1742Ser Val Pro Phe Asp Ile Arg be Ile Leu Asp Asp will Leu Ser Leu

545 550 555545 550 555

ttc tet gag gag tca agg aac aaa ggc att gag ctc gcg gtt ttc gtt 1790Phe Ser Glu Glu Ser Arg Asn Lys Gly Ile Glu Leu Ala Val Phe Val560 565 570ttc tet gag gag tca agg aac aaa ggc att gag ctc gcg gtt ttc gtt 1790Phe Ser Glu Glu Ser Arg Asn Lys Gly Ile Glu Leu Ala Val Phe Val560 565 570

tca gac aaa gta cca gag ata gtc aaa gga gat tca ggg aga ttt aga 1838Ser Asp Lys vai Pro Glu Ile vai Lys Gly Asp Ser Gly Arg Phe Arg575 580 585 590gca aaa gta aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 1838Ser Asp Lys Goes To Glu Ile Goes Lys Gly Asp Being Gly Arg Phe Arg575 580 585 590

cag ata ate ata aac ctt gtt gga aat tcg gtt aaa ttc aca gag aaa 1886Gln Ile Ile Ile Asn Leu vai Gly Asn Ser Val Lys Phe Thr Glu Lyscag ata till ata aac ctt gtt gga aat tcg gtt aaa ttc aca gag aaa 1886Gln Ile Ile Ile Asn Leu goes Gly Asn Ser Val Lys Phe Thr Glu Lys

595 600 605595 600 605

gga cat ate ttt gtt aaa gtc cat ctt gcg gaa caa tca aaa gat gaa 1934Gly His Ile Phe Val Lys Val His Leu Ala Glu Gln Ser Lys Asp Glu610 615 620gga cat till ttt gtt aaa gtc cat ctt gcg gaa caa tca aaa gat gaa 1934Gly His Ile Phe Val Lys Val His Leu Ala Glu Gln Ser Lys Asp Glu610 615 620

tet gaa ccg aaa aat gea ttg aat ggt gga gtg tet gaa gaa atg ate 1982Ser Glu Pro Lys Asn Ala Leu Asn Gly Gly Val Ser Glu Glu Met Iletet gaa ccg aaa aat gea ttg aat ggt gga gtg tet gaa gaa atg until 1982Ser Glu Pro Lys Asn Ala Leu Asn Gly Gly Val Ser Glu Glu Met Ile

625 630 635625 630 635

gtt gtt tcc aaa cag tca agt tac aac aca ttg age ggt tac gaa gct 2030Val Val Ser Lys Gln Ser Ser Tyr Asn Thr Leu Ser Gly Tyr Glu Ala640 645 650gtt gtt tcc aaa cag tca agt tac aac aca ttg age ggt tac gaa gct 2030Val Val Ser Lys Gln Ser Ser Tyr Asn Thr Leu Ser Ser Gly Tyr Glu Ala640 645 650

gct gat ggt cgg aat age tgg gat tca ttc aag cat ttg gtc tet gag 2078Ala Asp Gly Arg Asn ser Trp Asp ser Phe Lys His Leu Val Ser Glu655 660 665 670gct gat ggt cgg aat age tgg gat tca ttc aag cat ttg gtc tet gag 2078Ala Asp Gly Arg Asn Be Trp Asp Be Phe Lys His Leu Val Ser Glu655 660 665 670

gag cag tca tta tcg gag ttt gat att tet age aat gtt agg ctt atg 2126Glu Gln Ser Leu Ser Glu Phe Asp Ile Ser Ser Asn Val Arg Leu Met675 680 685gtt tca ate gaa gac acg ggt att gga ate cct tta gtt gea caa ggc 2174vai Ser Ile Glu Asp Thr Gly Ile Gly Ile Pro Leu Val Ala Gln Glygag cag tca tta tcg gag ttt gat att tet age aat gtt agg ctt atg 2126Glu Gln Ser Leu Ser Glu Phe Asp Ile Ser Ser Asn Val Arg Leu Met675 680 685gtt tca till gaa gac acg ggt att gga till cct tta gtt gea caa ggc 2174vai Ser Ile Glu Asp Thr Gly Ile Gly Ile Pro Read Val Wing Gln Gly

690 695 700690 695 700

cgt gtg ttt atg ccg ttt atg caa gea gat age tcg act tca aga aac 2222Arg Val Phe Met Pro Phe Met Gln Ala Asp Ser Ser Thr Ser Arg Asn705 710 715cgt gtg ttt atg ccg ttt atg caa gea gat age tcg act tca aga aac 2222Arg Val Phe Met Pro Phe Met Gln Asp Ser Ser Thr Thr Asn705 710 715

tat gga ggt act ggt att ggt ttg agt ata age aag tgt ctt gtt gaa 2270Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile Ser Lys Cys Leu Val Glu720 725 730tat gga ggt act ggt att ggt ttg agt ata age aag tgt ctt gtt gaa 2270Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile Ser Lys Cys Leu Val Glu720 725 730

ctt atg cgt ggt cag ata aat ttc ata age cgg cct cat att gga age 2318Leu Met Arg Gly Gln Ile Asn Phe Ile Ser Arg Pro His Ile Gly Ser735 740 745 750ctt atg cgt ggt cag ata aat ttc ata age cgg cct cat att gga age 2318Leu Met Arg Gly Gln Ile Asn Phe Ile Ser Arg Pro His Ile Gly Ser735 740 745 750

acg ttc tgg ttc acg gct gtt tta gag aaa tgc gat aaa tgc agt gcg 2366Thr Phe Trp Phe Thr Ala vai Leu Glu Lys Cys Asp Lys Cys Ser Ala 755 760 765acg ttc tgg ttc acg gct gtt tta gag aaa tgc gat aaa tgc agt gcg 2366Thr Phe Trp Phe Thr Wing goes Leu Glu Lys Cys Asp Lys Cys Ser Ala 755 760 765

att aac cat atg aag aaa cct aat gtg gaa cac ttg cct tet act ttt 2414Ile Asn His Met Lys Lys Pro Asn vai Glu His Leu Pro Ser Thr Pheatt aac cat atg aag aaa cct aat gtg gaa cac ttg cct tet act ttt 2414Ile Asn His Met Lys Lys Pro Asn Go Glu His Leu Pro Ser Thr Phe

770 775 780770 775 780

aaa gga atg aaa gct ata gtt gtt gat gct aag cct gtt aga gct gct 2462Lys Gly Met Lys Ala Ile vai vai Asp Ala Lys Pro vai Arg Ala Ala785 790 795aaa gga atg aaa gct ata gtt gtt gat gct aag cct gtt aga gct gct 2462Lys Gly Met Lys Ala Ile Go Go Asp Ala Lys Pro Go Arg Ala Ala785 790 795

gtg act aga tac cat atg aaa aga ctc gga ate aat gtt gat gtc gtg 2510Val Thr Arg Tyr His Met Lys Arg Leu Gly Ile Asn Val Asp Val Val800 805 810gtg act aga tac cat atg aaa aga ctc gga until aat gtt gat gtc gtg 2510Val Thr Arg Tyr His Met Lys Arg Leu Gly Ile Asn Val Asp Val Val800 805 810

aca agt ctc aaa acc gct gtt gtt gea gct gct gcg ttt gaa aga aac 2558Thr Ser Leu Lys Thr Ala vai vai Ala Ala Ala Ala Phe Glu Arg Asn815 820 825 830aca agt ctc aaa acc gct gtt gtt gea gct gct gcg ttt gaa aga aac 2558Thr Be Leu Lys Thr Wing go go Wing Wing Wing Wing Phe Glu Arg Asn815 820 825 830

ggt tet cct ctc cca aca aaa ccg caa ctt gat atg ate tta gta gag 2606Gly Ser Pro Leu Pro Thr Lys Pro Gln Leu Asp Met Ile Leu vai Glu 835 840 845ggt tet cct ctc aca aaa ccg caa ctt gat atg till tta gta gag 2606Gly Ser Pro Leu Pro Thr Lys Pro Gln Leu Asp Met Ile Leu Go Glu 835 840 845

aaa gat tca tgg att tca act gaa gat aat gac tca gag att cgt tta 2654Lys Asp Ser Trp Ile Ser Thr Glu Asp Asn Asp Ser Glu Ile Arg Leu850 855 860aaa gat tca tgg att tca act gaa gat aat gac tca gag att cgt tta 2654Lys Asp Be Trp Ile Be Thr Glu Asp Asn Asp Be Glu Ile Arg Leu850 855 860

ttg aat tca aga acc aac gga aac gtt cat cac aag tct ccg aaa cta 2702Leu Asn Ser Arg Thr Asn Gly Asn vai His His Lys Ser Pro Lys Leu865 870 875ttg aat tca aga acc aac gga aac gtt cat cac aag tct ccg aaa cta 2702Leu Asn Ser Arg Thr Asn Gly Asn Goes His His Lys Ser Pro Lys Leu865 870 875

gct cta ttc gca aca aac ate aca aat tcg gag ttc gac aga gct aaa 2750Ala Leu Phe Ala Thr Asn Ile Thr Asn ser Glu Phe Asp Arg Ala Lysgct cta ttc gca aca aac until aca aat tcg gag ttc gac aga gct aaa 2750Ala Leu Phe Ala Thr Asn Ile Thr Asn be Glu Phe Asp Arg Ala Lys

880 885 890880 885 890

tcc gca gga ttt gca gat acg gta ata atg aaa ccg tta aga gca age 2798Ser Ala Gly Phe Ala Asp Thr vai Ile Met Lys Pro Leu Arg Ala Ser895 900 905 910tcc gca gga ttt gca gat acg gta ata atg aaa ccg tta aga gca age 2798Ser Wing Gly Phe Wing Asp Thr will Ile Met Lys Pro Leu Arg Wing Ser895 900 905 910

atg att ggg gcg tgt ctg caa caa gtt ctc gag ctg aga aaa aca aga 2846Met Ile Gly Ala Cys Leu Gln Gln vai Leu Glu Leu Arg Lys Thr Argatg att ggg gcg tgt ctg caa caa gtt ctc gag ctg aga aaa aca aga 2846Met Ile Gly Ala Cys Leu Gln Gln Go Leu Glu Leu Arg Lys Thr Arg

915 920 925915 920 925

caa caa cat cca gaa gga tca tca ccc gca act ctc aag age ttg ctt 2894Gln Gln His Pro Glu Gly Ser Ser Pro Ala Thr Leu Lys Ser Leu Leu930 935 940caa caa cat cca gaa gga tca tca ccc gca act ctc aag age ttg ctt 2894Gln Gln His Pro Glu Gly Ser Sera Ala Thr Leu Lys Ser Leu Leu930 935 940

aca ggg aag aag att ctt gtg gtt gat gat aat ata gtt aac agg aga 2942Thr Gly Lys Lys Ile Leu vai Val Asp Asp Asn Ile Val Asn Arg Arg945 950 955ggg aag aag att ctt gtg gtt gat gat aat ata gtt aac agg aga 2942Thr Gly Lys Lys Ile Leu goes Val Asp Asp Asn Ile Val Asn Arg Arg945 950 955

gta gct gca gga gct ctc aag aaa ttt gga gca gaa gtg gtt tgt gca 2990Val Ala Ala Gly Ala Leu Lys Lys Phe Gly Ala Glu Val vai Cys Alagta gct gca gga gct ctc aag aaa ttt gga gca gaa gtg gtt tgt gca 2990Val Wing Wing Gly Wing Leu Lys Lys Phe Gly Wing Glu Val goes Cys Wing

960 965 970960 965 970

gag agt ggt caa gtt gct ttg ggt ttg ctt cag att cca cac act ttc 3038Glu Ser Gly Gln Val Ala Leu Gly Leu Leu Gln Ile Pro His Thr Phe975 980 985 990gag agt ggt caa gtt gct ttg ggt ttg ctt cag att cca cac act ttc 3038Glu Ser Gly Gln Val Ala Leu Gly Leu Leu Gln Ile Pro His Thr Phe975 980 985 990

gat gct tgc ttc atg gat att caa atg cca cag atg gac gga ttt gaa 3086Asp Ala Cys Phe Met Asp Ile Gln Met Pro Gln Met Asp Gly Phe Glugat gct tgc ttc atg gat att caa atg cca cag atg gac gga ttt gaa 3086Asp Cys Wing Phe Met Asp Ile Gln Met Pro Gln Met Asp Gly Phe Glu

995 1000 1005995 1000 1005

gca act cgt cag ata aga atg atg gag aag gaa gct aaa gag aag 3131Ala Thr Arg Gln lie Arg Met Met Glu Lys Glu Ala Lys Glu Lys1010 1015 1020gca act cgt cag ata ag atg atg gag aag gaa gct aaa gag aag 3131Ala Thr Arg Gln Lie Arg Met Met Glu Lys Glu Ala Lys Glu Lys1010 1015 1020

acg aat ctc gaa tgg cat tta ccg att cta gcg atg act gcg gat 3176Thr Asn Leu Glu Trp His Leu Pro lie Leu Ala Met Thr Ala Aspacg aat ctc gaa tgg cat tta ccg att cta gcg atg act gcg gat 3176Thr Asn Leu Glu Trp His Leu Pro lie Leu Ala Met Thr Ala Asp

1025 1030 10351025 1030 1035

gtg ata cac gcg acc tac gag gaa tgt ctg aaa agt ggg atg gat 3221vai Ile His Ala Thr Tyr Glu Glu Cys Leu Lys Ser Gly Met Asp 1040 1045 1050gtg ata cac gcg acc tac gag gaa tgt ctg aaa agt ggg atg gat 3221vai Ile His Wing Thr Tyr Glu Glu Cys Leu Lys Ser Gly Met Asp 1040 1045 1050

ggt tac gtc tcc aaa cct ttt gaa gaa gag aat ctc tat aaa tcc 3266Gly Tyr Val Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr Lys Serggt tac gtc tcc aaa cct ttt gaa gaa gag aat ctc tat aaa tcc 3266Gly Tyr Val Lys Pro Phe Glu Glu Glu Asn Leu Tyr Lys Ser

1055 1060 10651055 1060 1065

gtt gcc aaa tca ttc aaa cct aat cct ate tca cct tcg tcg taa 3311Val Ala Lys ser Phe Lys Pro Asn Pro Ile Ser Pro Ser Ser1070 1075 1080gtt gcc aaa tca ttc aaa cct aat cct until tca cct tcg tcg taa 3311Val Ala Lys be Phe Lys Pro Asn Pro Ile Ser Pro Ser1070 1075 1080

tccaatcttc cggcgagttt ttttctctcc gcagccggaa gaatggactg cttctgctga 3371ttgattggga taaatatgca ttttggtttc tgtacatata gtaggttcac aatetagaga 3431ttttgaaggt tttttttaat ttctcactta agtaatgtag cttgccatga ctagtgtatg 3491ttgttaaacg acgacgtcta agatggttca gtgttgatct tagcgtaagt attaatccca 3551tgggaatcgg ttgtactgta tcagatttgg ttagtcgttt aancattgta atgttctaat 3611aatcactttt tcatatatan cctcatgtta taacatgaga cgagaccatt ttgattaaaa 3671aaaaaaaaaa aaaaaaaaaa aaa 3694tccaatcttc cggcgagttt ttttctctcc gcagccggaa gaatggactg cttctgctga 3371ttgattggga taaatatgca ttttggtttc tgtacatata gtaggttcac aatetagaga 3431ttttgaaggt tttttttaat ttctcactta agtaatgtag cttgccatga ctagtgtatg 3491ttgttaaacg acgacgtcta agatggttca gtgttgatct tagcgtaagt attaatccca 3551tgggaatcgg ttgtactgta tcagatttgg ttagtcgttt aancattgta atgttctaat 3611aatcactttt tcatatatan cctcatgtta taacatgaga cgagaccatt ttgattaaaa 3671aaaaaaaaaa aaaaaaaaaa aaa 3694

<210> 9<211> 1080<212> PRT<210> 9 <211> 1080 <212> PRT

<213> Arabidopsis thaliana<400> 9<213> Arabidopsis thaliana <400> 9

Met Arg Arg Asp Phe Val Tyr Asn Asn Asn Ala Met Phe Asn Pro Leu1 5 10 15Met Arg Arg Asp Phe Val Tyr Asn Asn Asn Wing Met Phe Asn Pro Leu1 5 10 15

Thr Thr His Tyr Ser Ser Asp Met Asn Trp Ala Leu Asn Asn His GlnThr Thr His Tyr Being Ser Asp Met Asn Trp Wing Read Asn Asn His Gln

20 25 3020 25 30

Glu Glu Glu Glu Glu Pro Arg Arg lie Glu Ile Ser Asp Ser Glu Ser35 40 45Glu Glu Glu Glu Glu Pro Arg Arg Lie Glu Ile Ser Asp Ser Glu Ser35 40 45

Leu Glu Asn Leu Lys Ser Ser Asp Phe Tyr Gln Leu Gly Gly Gly Gly50 55 60Read Glu Asn Read Le Lys Be Ser Asp Phe Tyr Gln Read Gly Gly Gly Gly50 55 60

Ala Leu Asn Ser Ser Glu Lys Pro Arg Lys Ile Asp Phe Trp Arg Ser65 70 75 80Wing Read Asn Be Glu Lys Pro Arg Lys Ile Asp Phe Trp Arg Ser65 70 75 80

Gly Leu Met Gly Phe Ala Lys Met Gln Gln Gln Gln Gln Leu Gln HisGly Leu Met Gly Phe Ala Lys Met Gln Gln Gln Gln Gln Glu Leu Gln His

85 90 9585 90 95

Ser Val Ala Val Lys Met Asn Asn Asn Asn Asn Asn Asp Leu Met Gly100 105 110Ser Val Wing Val Lys Met Asn Asn Asn Asn Asn Asn Asn Asp Leu Met Gly100 105 110

Asn Lys Lys Gly Ser Thr Phe Ile Gln Glu His Arg Ala Leu Leu ProAsn Lys Lys Gly Be Thr Phe Ile Gln Glu His Arg Wing Read Leu Pro

115 120 125115 120 125

Lys Ala Leu Ile Leu Trp Ile Ile Ile Val Gly Phe Ile Ser Ser Gly130 135 140Lys Wing Leu Ile Leu Trp Ile Ile Ile Val Gly Phe Ile Ser Ser Gly130 135 140

Ile Tyr Gln Trp Met Asp Asp Ala Asn Lys Ile Arg Arg Glu Glu vai145 150 155 160Ile Tyr Gln Trp Met Asp Asp Wing Asn Lys Ile Arg Arg Glu Glu vai145 150 155 160

Leu Val Ser Met Cys Asp Gln Arg Ala Arg Met Leu Gln Asp Gln PheLeu Val Ser Met Cys Asp Gln Arg Wing Arg Met Leu Gln Asp Gln Phe

165 170 175165 170 175

Ser vai Ser vai Asn His Val His Ala Leu Ala Ile Leu vai Ser Thr180 185 190Be will Be will Asn His Val His Wing Leu Wing Ile Leu will Be Thr180 185 190

Phe His Tyr His Lys Asn Pro Ser Ala Ile Asp Gln Glu Thr Phe AlaPhe His Tyr His Lys Asn Pro Be Wing Ile Asp Gln Glu Thr Phe Wing

195 200 205195 200 205

Glu Tyr Thr Ala Arg Thr Ala Phe Glu Arg Pro Leu Leu Ser Gly vai210 215 220Glu Tyr Thr Wing Arg Thr Wing Phe Glu Arg Pro Read Leu Ser Gly vai210 215 220

Ala Tyr Ala Glu Lys Val Val Asn Phe Glu Arg Glu Met Phe Glu Arg225 230 235 240Tyr Wing Glu Wing Lys Val Val Asn Phe Glu Arg Glu Met Phe Glu Arg225 230 235 240

Gln His Asn Trp Val Ile Lys Thr Met Asp Arg Gly Glu Pro Ser ProGln His Asn Trp Val Ile Lys Thr Met Asp Arg Gly Glu Pro Ser Pro

245 250 255245 250 255

vai Arg Asp Glu Tyr Ala Pro Val Ile Phe Ser Gln Asp Ser vai Ser260 265 270Go Arg Asp Glu Tyr Ala Pro Val Ile Phe Ser Go Gln Asp Ser Go Ser260 265 270

Tyr Leu Glu Ser Leu Asp Met Met Ser Gly Glu Glu Asp Arg Glu AsnTyr Read Glu Be Read Asp Met Met Be Gly Glu Asp Arg Glu Asn

275 280 285275 280 285

Ile Leu Arg Ala Arg Glu Thr Gly Lys Ala Val Leu Thr Ser Pro Phe290 295 300Ile Leu Arg Wing Arg Glu Thr Gly Lys Wing Val Leu Thr Be Pro Phe290 295 300

Arg Leu Leu Glu Thr His His Leu Gly vai Val Leu Thr Phe Pro Val305 310 315 320Arg Leu Leu Glu Thr His His Leu Gly Goes Val Leu Thr Phe Pro Val305 310 315 320

Tyr Lys Ser Ser Leu Pro Glu Asn Pro Thr Val Glu Glu Arg Ile Ala4 325 330 335Tyr Lys Ser Serve Leu Pro Glu Asn Pro Thr Val Glu Glu Arg Ile Ala4 325 330 335

Ala Thr Ala Gly Tyr Leu Gly Gly Ala Phe Asp Val Glu Ser Leu ValWing Thr Wing Wing Gly Tyr Leu Gly Gly Wing Phe Asp Val Glu Ser Leu Val

340 345 350340 345 350

Glu Asn Leu Phe Gly Gln Leu Ala Gly Asn Gln Ala Ile Val Val His 355 360 365Glu Asn Leu Phe Gly Gln Leu Wing Gly Asn Gln Wing Ile Val Val His 355 360 365

Val Tyr Asp lie Thr Asn Ala Ser Asp Pro Leu vai Met Tyr Gly AsnVal Tyr Asp lie Thr Asn Ala Ser Asp Pro Leu goes Met Tyr Gly Asn

370 375 380370 375 380

Gln Asp Glu Glu Ala Asp Arg ser Leu Ser His Glu Ser Lys Leu Asp385 390 395 400Gln Asp Glu Glu Wing Asp Arg Be Read His Be Glu Be Lys Leu Asp385 390 395 400

Phe Gly Asp Pro Phe Arg Lys His Lys Met Il e Cys Arg Tyr His GlnPhe Gly Asp Pro Phe Arg Lys His Lys Met Il and Cys Arg Tyr His Gln

405 410 415405 410 415

Lys Ala Pro Ile Pro Leu Asn Val Leu Thr Thr vai Pro Leu Phe PheLys Wing Pro Ile Pro Leu Asn Val Leu Thr Thr Go Pro Leu Phe Phe

420 425 430420 425 430

Ala Ile Gly Phe Leu vai Gly Tyr Ile Leu Tyr Gly Ala Ala Met His 435 440 445Wing Ile Gly Phe Leu Goes Gly Tyr Ile Leu Tyr Gly Wing Wing Met His 435 440 445

Ile vai Lys vai Glu Asp Asp Phe His Glu Met Gln Glu Leu Lys vaiIle will Lys will Glu Asp Asp Phe His Glu Met Gln Glu Leu Lys will

450 455 460450 455 460

Arg Ala Glu Ala Ala Asp Val Ala Lys Ser Gln Phe Leu Ala Thr Val465 470 475 480Arg Wing Glu Wing Wing Asp Val Wing Lys Ser Gln Phe Leu Wing Thr Val465 470 475 480

Ser His Glu Ile Arg Thr Pro Met Asn Gly Ile Leu Gly Met Leu AlaBe His Glu Ile Arg Thr Pro Met Asn Gly Ile Leu Gly Met Leu Ala

485 490 495485 490 495

Met Leu Leu Asp Thr Glu Leu Ser Ser Thr Gln Arg Asp Tyr Ala GlnMet Leu Read Asp Thr Glu Read Be Ser Thr Gln Arg Asp Tyr Wing Gln

500 505 510500 505 510

Thr Ala Gln Val Cys Gly Lys Ala Leu Ile Ala Leu Ile Asn Glu Val 515 520 525Thr Wing Gln Val Cys Gly Lys Wing Leu Ile Wing Leu Ile Asn Glu Val 515 520 525

Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys Leu Glu Leu Glu Ser vaiLeu Asp Arg Wing Lys Ile Glu Wing Gly Lys Leu Glu Leu Glu Be going

530 535 540530 535 540

Pro Phe Asp Ile Arg ser Ile Leu Asp Asp Val Leu Ser Leu Phe Ser545 550 555 560Pro Phe Asp Ile Arg Be Ile Leu Asp Asp Val Leu Be Leu Phe Ser545 550 555 560

Glu Glu Ser Arg Asn Lys Gly lie Glu Leu Ala Val Phe vai Ser AspGlu Glu Be Arg Asn Lys Gly lie Glu Leu Wing Val Phe Will Be Asp

565 570 575565 570 575

Lys Val Pro Glu Il e vai Lys Gly Asp Ser Gly Arg Phe Arg Gln Ile580 585 590Lys Val Pro Glu Il And Go Lys Gly Asp Ser Gly Arg Phe Arg Gln Ile580 585 590

Ile Ile Asn Leu vai Gly Asn ser Val Lys Phe Thr Glu Lys Gly HisIle Ile Asn Leu will Gly Asn be Val Lys Phe Thr Glu Lys Gly His

595 600 605595 600 605

Ile Phe Val Lys vai His Leu Ala Glu Gln Ser Lys Asp Glu Ser Glu610 615 620Ile Phe Val Lys Goes His Leu Wing Glu Gln Ser Lys Asp Glu Ser Glu610 615 620

Pro Lys Asn Ala Leu Asn Gly Gly Val ser Glu Glu Met Ile vai Val625 630 635 640Pro Lys Asn Wing Read Asn Gly Gly Val Be Glu Glu Met Ile Go Val625 630 635 640

Ser Lys Gln Ser Ser Tyr Asn Thr Leu Ser Gly Tyr Glu Ala Ala Asp645 650 655Ser Lys Gln Ser Ser Tyr Asn Thr Read Ser Ser Gly Tyr Glu Ala Wing Asp645 650 655

Gly Arg Asn ser Trp Asp Ser Phe Lys His Leu Val Ser Glu Glu Gln660 665 670Gly Arg Asn Be Trp Asp Be Phe Lys His Leu Val Be Glu Glu Gln660 665 670

Ser Leu Ser Glu Phe Asp Ile Ser ser Asn Val Arg Leu Met Val SerSer Leu Ser Glu Phe Asp Ile Ser Ser Asn Val Arg Ser Leu Met Val Ser

675 680 685675 680 685

Ile Glu Asp Thr Gly Ile Gly Ile Pro Leu Val Ala Gln Gly Arg vai690 695 700Ile Glu Asp Thr Gly Ile Gly Ile Pro Read Val Wing Gln Gly Arg vai690 695 700

Phe Met Pro Phe Met Gln Ala Asp Ser ser Thr Ser Arg Asn Tyr Gly705 710 715 720Phe Met Pro Phe Met Gln Wing Asp Be Be Thr Be Arg Asn Tyr Gly705 710 715 720

Gly Thr Gly Ile Gly Leu Ser Ile Ser Lys Cys Leu Val Glu Leu Met725 730 735Gly Thr Gly Ile Gly Ile Ile Ile Be Lys Cys Leu Val Glu Leu Met725 730 735

Arg Gly Gln Ile Asn Phe Ile Ser Arg Pro His Ile Gly Ser Thr Phe740 745 750Arg Gly Gln Ile Asn Phe Ile Be Arg Pro His Ile Gly Be Thr Phe740 745 750

Trp Phe Thr Ala vai Leu Glu Lys Cys Asp Lys Cys Ser Ala Ile AsnTrp Phe Thr Wing Go Read Glu Lys Cys Asp Lys Cys Be Wing Ile Asn

755 760 765755 760 765

His Met Lys Lys Pro Asn vai Glu His Leu Pro Ser Thr Phe Lys Gly770 775 780His Met Lys Lys Pro Asn Goes Glu His Leu Pro Be Thr Phe Lys Gly770 775 780

Met Lys Ala Ile Val Val Asp Ala Lys Pro vai Arg Ala Ala Val Thr785 790 795 800Met Lys Wing Ile Val Val Asp Wing Lys Pro Goes Arg Wing Wing Val Thr785 790 795 800

Arg Tyr His Met Lys Arg Leu Gly Ile Asn Val Asp Val Val Thr Ser805 810 815Arg Tyr His Met Lys Arg Read Gly Ile Asn Val Asp Val Val Thr Ser805 810 815

Leu Lys Thr Ala Val Val Ala Ala Ala Ala Phe Glu Arg Asn Gly Ser820 825 830Leu Lys Thr Wing Val Val Wing Wing Wing Wing Wing Phe Wing Glu Arg Asn Gly Ser820 825 830

Pro Leu Pro Thr Lys Pro Gln Leu Asp Met Ile Leu Val Glu Lys Asp835 840 845Pro Read Pro Thr Lys Pro Gln Read Asp Met Ile Read Val Glu Lys Asp835 840 845

Ser Trp lie Ser Thr Glu Asp Asn Asp Ser Glu Ile Arg Leu Leu AsnSer Trp lie Ser Thr Glu Asp Asn Asp Ser Glu Ile Arg Leu Leu Asn

850 855 860850 855 860

Ser Arg Thr Asn Gly Asn Val His His Lys Ser Pro Lys Leu Ala Leu865 870 875 880Be Arg Thr Asn Gly Asn Val His His Lys Be Pro Lys Leu Wing Leu865 870 875 880

Phe Ala Thr Asn Ile Thr Asn Ser Glu Phe Asp Arg Ala Lys Ser AlaPhe Ala Thr Asn Ile Thr Asn Be Glu Phe Asp Arg Ala Lys Be Ala

885 890 895885 890 895

Gly Phe Ala Asp Thr Val Ile Met Lys Pro Leu Arg Ala ser Met Ile900 905 910Gly Phe Wing Asp Thr Val Ile Met Lys Pro Read Arg Wing Be Met Ile900 905 910

Gly Ala Cys Leu Gln Gln Val Leu Glu Leu Arg Lys Thr Arg Gln Gln915 920 925Gly Wing Cys Leu Gln Gln Val Leu Glu Leu Arg Lys Thr Arg Gln915 920 925

His Pro Glu Gly Ser Ser Pro Ala Thr Leu Lys Ser Leu Leu Thr GlyHis Pro Glu Gly Be Be Pro Wing Thr Read Lys Be Read Leu Thr Gly

930 935 940930 935 940

Lys Lys Ile Leu Val Val Asp Asp Asn Ile Val Asn Arg Arg vai Ala945 950 955 960Lys Lys Ile Leu Val Val Asp Asp Asn Ile Val Asn Arg Arg goes Ala945 950 955 960

Ala Gly Ala Leu Lys Lys Phe Gly Ala Glu vai vai Cys Ala Glu SerGly Wing Wing Read Lys Lys Phe Gly Wing Glu Go Go Cys Wing Glu Ser

965 970 975965 970 975

Gly Gln vai Ala Leu Gly Leu Leu Gln Ile Pro His Thr Phe Asp Ala980 985 990Gly Gln goes Ala Leu Gly Leu Leu Gln Ile Pro His Thr Phe Asp Ala980 985 990

Cys Phe Met Asp Ile Gln Met Pro Gln Met Asp Gly Phe Glu Ala Thr995 1000 1005Cys Phe Met Asp Ile Gln Met Pro Gln Met Asp Gly Phe Glu Wing Thr995 1000 1005

Arg Gln Ile Arg Met Met Glu Lys Glu Ala Lys Glu Lys Thr AsnArg Gln Ile Arg Met Met Glu Lys Glu Wing Lys Glu Lys Thr Asn

1010 1015 10201010 1015 1020

Leu Glu Trp His Leu Pro Ile Leu Ala Met Thr Ala Asp vai Ile1025 1030 1035Leu Glu Trp His Leu Pro Ile Leu Wing Met Thr Wing Asp goes Ile1025 1030 1035

His Ala Thr Tyr Glu Glu Cys Leu Lys Ser Gly Met Asp Gly TyrHis Wing Thr Tyr Glu Glu Cys Read Lys Ser Gly Met Asp Gly Tyr

1040 1045 10501040 1045 1050

Val Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr Lys Ser Val Ala1055 1060 1065Val Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr Lys Ser Val Ala1055 1060 1065

Lys ser Phe Lys Pro Asn Pro Ile Ser Pro Ser Ser1070 1075 1080Lys to be Phe Lys to be

<210> 10<211> 5474<210> 10 <211> 5474

<212> DNA<212> DNA

<213> Oryza (rice)<220><213> Oryza (rice) <220>

<221> exon<221> exon

<222> (1)..(348)<220><222> (1) .. (348) <220>

<221> Intron<221> Intron

<222> (349)..(711)<220><222> (349) .. (711) <220>

<221> exon<221> exon

<222> (712)..(945)<222> (712) .. (945)

<220><220>

<221> Intron<221> Intron

<222> (946)..(1040)<220><222> (946) .. (1040) <220>

<221> exon<221> exon

<222> (1041)..(1207)<220><222> (1041) .. (1207) <220>

<221> Intron<221> Intron

<222> (1208)..(1508)<220><222> (1208) .. (1508) <220>

<221> exon<221> exon

<222> (1509)..(1727)<220><222> (1509) .. (1727) <220>

<221> Intron<221> Intron

<222> (1728)..(1818)<220><222> (1728) .. (1818) <220>

<221> exon<221> exon

<222> (1819)..(2005)<220><222> (1819) .. (2005) <220>

<221> Intron<222> (2006)..(2087)<220><221> Intron <222> (2006) .. (2087) <220>

<221> exon<222> (2088)..(2139)<220><221> exon <222> (2088) .. (2139) <220>

<221> Intron<222> (2140)..(2233)<220><221> Intron <222> (2140) .. (2233) <220>

<221> exon<222> (2234)..(2463)<220><221> exon <222> (2234) .. (2463) <220>

<221> Intron<222> (2464)..(2694)<220><221> exon<221> Intron <222> (2464) .. (2694) <220> <221> exon

<222> (2695)..(2793)<220><222> (2695) .. (2793) <220>

<221> Intron<222> (2794)..(2910)<220><221> Intron <222> (2794) .. (2910) <220>

<221> exon<221> exon

<222> (2911)..(3641)<222> (2911) .. (3641)

<220><220>

<221> Intron<222> (3642)..(4079)<220><221> Intron <222> (3642) .. (4079) <220>

<221> exon<222> (4080)..(4595)<220><221> Intron<221> exon <222> (4080) .. (4595) <220> <221> Intron

<222> (4596)..(5215)<220><221> exon<222> (4596) .. (5215) <220> <221> exon

<222> (5216)..(5474)<222> (5216) .. (5474)

<400> 10<400> 10

atg ggt gtg gga gga ggc gga gga gga gga gga ggg gag gcg gcg gcg 48Met Gly Val Gly Gly Gly Gly Gly Gly Gly Gly Gly Glu Ala Ala Ala1 5 10 15atg ggt gtg gga gga ggg gga gga gga gga gga gga ggg gag gcg gcg gcg 48Met Gly Val Gly Gly Gly Gly Gly Gly Gly Gly Gly

gcg gtg gcg gtg gag ggg gat gag gcg ggg aag ggg agg agg tgg tgg 96gcg gtg gcg gtg gag ggg gat gag gcg ggg aag ggg agg agg tgg tgg 96

Ala vai Ala Val Glu Gly Asp Glu Ala Gly Lys Gly Arg Arg Trp Trp20 25 30Wing goes Wing Val Glu Gly Asp Glu Wing Gly Lys Gly Arg Arg Trp Trp20 25 30

agg gtg aag gtg aag ctg age acg gtg gcg gtg gtg gcg tgg gtg ctg 144Arg vai Lys vai Lys Leu Ser Thr Val Ala Val Val Ala Trp Val Leu35 40 45agg gtg aag gtg aag ctg age acg gtg gcg gtg gtg gtg gtg gtg ctg 144Arg will Lys will Lys Leu Ser Thr Val Wing Val Val Wing Val Tru Val Leu35 40 45

gcg tcg gcg gcg ctc tgg gcg ggg ctg cac tgg cgc ttc cgc cgc gcg 192Ala Ser Ala Ala Leu Trp Ala Gly Leu His Trp Arg Phe Arg Arg Ala50 55 60gcg tcg gcg gcg ctc tgg gcg ggg ctg cac tgg cgc ttc cgc cgc gcg 192Ala Ser Ala Ala Leu Trp Ala Gly Leu His Trp Arg Phe Arg Arg Ala50 55 60

gcg ctg cac aag gcc gag gag gcc ctc gtc tgc atg tgc gag gag cgc 240Ala Leu His Lys Ala Glu Glu Ala Leu Val Cys Met Cys Glu Glu Arg65 70 75 80gcg ctg cac aag gcc gag gag gcc ctc gtc tgc atg tgc gag gag cgc 240Ala Leu His Lys Wing Glu Glu Wing Leu Val Cys Met Cys Glu Glu Arg65 70 75 80

gcc cgc atg ctg cag gac cag ttc gcc gtc tcc gtc aac cac gtc cac 288Ala Arg Met Leu Gln Asp Gln Phe Ala vai Ser Val Asn His Val His85 90 95gcc cgc atg ctg cag gac cag ttc gcc gtc tcc gtc aac cac gtc cac 288Ala Arg Met Leu Gln Asp Gln Phe Ala Will Be Val Asn His Val His85 90 95

gcc ctc gcc ate ctc gtc gcc acc ttc cac tac gac aag cac cct ccc 336Ala Leu Ala Ile Leu Val Ala Thr Phe His Tyr Asp Lys His Pro Pro100 105 110gcc ctc gcc up to ctc gtc gcc acc ttc cac tac gac aag cac cct ccc 336Ala Leu Ala Ile Leu Val Ala Thr Phe His Tyr Asp Lys His Pro Pro 105 110 110

gcc ctc gac cag gtcggcccga actccgacga gctcttccgc cgccgccgcg 388Ala Leu Asp Gln115gcc ctc gac cag gtcggcccga actccgacga gctcttccgc cgccgccgcg 388Ala Leu Asp Gln115

atgatcctgt tgcatctgtt gtttttttgc cccccgcggt taattgcgat aatgcctcga 448tttttactcc acatcttgcc cgtgtacttc gctctgctgc ttcttcggct tcatttaatt 508ctaccgtgac cttccgtgtc agccatggaa gccatggatt actgttgctg tctcttgcta 568ttatatggag cgcacttttt gttgggaggg agtgaattga ttgtgcttgc ttgcttctgt 628tggtagtagt actagtgatt tctttggctg tgtggctgat gaatcttctt cgatgtgttg 688tgcgtgcgtg cgcgtgtgtg cag gac acg ttc gcc gtg tac gcc gcg agg acg 741atgatcctgt tgcatctgtt gtttttttgc cccccgcggt taattgcgat aatgcctcga 448tttttactcc acatcttgcc cgtgtacttc gctctgctgc ttcttcggct tcatttaatt 508ctaccgtgac cttccgtgtc agccatggaa gccatggatt actgttgctg tctcttgcta 568ttatatggag cgcacttttt gttgggaggg agtgaattga ttgtgcttgc ttgcttctgt 628tggtagtagt actagtgatt tctttggctg tgtggctgat gaatcttctt cgatgtgttg 688tgcgtgcgtg cgcgtgtgtg cag gac gtg acg ttc gcc gcc gcg tac agg acg 741

Asp Thr Phe Ala vai Tyr Ala Ala Arg Thr120 125Asp Thr Phe Wing Go Tyr Wing Wing Arg Thr120 125

tcc ttc gag cgg ccg ctg ctg age ggc gtg gcg tac gcg cag cgg gtg 789Ser Phe Glu Arg Pro Leu Leu Ser Gly Val Ala Tyr Ala Gln Arg vai130 135 140tcc ttc gag cgg ccg ctg ctg age ggc gtg gcg tac gcg cag cgg gtg 789Ser Phe Glu Arg Pro Read Leu Ser Gly Val Ala Tyr Ala Gln Arg vai130 135 140

gtg cac gcc gac agg gag age ttc gag cgg cag cag ggg tgg ate ate 837gtg cac gcc gac agg gag age ttc gag cgg cag cag ggg tgg until 837

Val His Ala Asp Arg Glu Ser Phe Glu Arg Gln Gln Gly Trp Ile Ile145 150 155Val His Wing Asp Arg Glu Be Phe Glu Arg Gln Gln Gly Trp Ile Ile145 150 155

aag acc atg aag cac gag ccg tcc ccg gcg cag gac gag tac gcc ccg 885Lys Thr Met Lys His Glu Pro Ser Pro Ala Gln Asp Glu Tyr Ala Proaag acc atg aag cac cc cc cc cc cc cc cc cc cc gag cc cc gag cc cc 885Lys Thr Met Lys His Glu Pro Ser Pro Ala Gln Asp Glu Tyr Ala Pro

160 165 170160 165 170

gtg ate tac tcc cag gag acc ate tcc tac ate gag ggc ctc gac gtc 933vai Ile Tyr Ser Gln Glu Thr Ile Ser Tyr Ile Glu Gly Leu Asp Val175 180 185 190gtg up to tac tcc cag gag acc up to tcc tac up to gag ggc ctc gac gtc 933vai Ile Tyr Ser Gln Glu Thr Ile Ser Tyr Ile Glu Gly Leu Asp Val175 180 185 190

atg tcc ggc gag gtgcgtttct tgggttacag cttcgcagct gctgctgcgg 985atg tcc ggc gag gtgcgtttct tgggttacag cttcgcagct gctgctgcgg 985

Met Ser Gly GluMet Ser Gly Glu

ttatcgccat gtccgctgct ctgaactgtg ctgctgggtg tgcttcggcc tgcag gag 1043Gluttatcgccat gtccgctgct ctgaactgtg ctgctgggtg tgcttcggcc tgcag gag 1043Glu

195195

gac agg gag aac ate ttg agg gcg agg gcg aca ggg aag gcc gtc ctc 1091Asp Arg Glu Asn Ile Leu Arg Ala Arg Ala Thr Gly Lys Ala vai Leu200 205 210gac agg gag aac to ttg agg ggg agg gcg aca ggg aag gcc gtc ctc 1091Asp Arg Glu Asn Ile Leu Arg Wing Arg Wing Thr Wing Gly Lys Wing goes Leu200 205 210 210

acg agg ccg ttc cgg ctg atg tcg aat cac ttg ggt gtt gtc ttg acg 1139Thr Arg Pro Phe Arg Leu Met Ser Asn His Leu Gly Val Val Leu Thracg agg ccg ttc cgg ctg atg tcg aat cac ttg ggt gtt gtc ttg acg 1139Thr Arg Pro Phe Arg Leu Met Ser Asn His Leu Gly Val Val Leu Thr

215 220 225215 220 225

ttt cct gtc tac ctc gtc gat ctc cca aat gat acc gcg gtg gag gat 1187Phe Pro Val Tyr Leu Val Asp Leu Pro Asn Asp Thr Ala Val Glu Asp 230 235 240ttt cct gtc tac ctc gtc gat ctc cca aat gat acc gcg gtg gag gat 1187Phe Pro Val Tyr Leu Val Asp Leu Pro Asn Thr Ala Val Glu Asp 230 235 240

cgt gtt gct gct act gea gg gtgagggatt actttacttt tctgaatgaa 1237cgt gtt gct gct act gea gg gtgagggatt actttacttt tctgaatgaa 1237

Arg Val Ala Ala Thr Ala Gly245Arg Val Wing Wing Thr Wing Wing Gly245

gattattctc tccaactgat tcctcttctg tctggaatcc actgccttca gctcttcgtt 1297ttgttgcagt cgttgttgga tgcttttagt agtggaaatg tgtgcgtttc agggatattt 1357gatcacatgc aacattttca ctcataactg gctgaaaaag ttttgcatta atagagctga 1417aatgtctaga tggataagca attgcagtgg tattttaagt acaacatgtg caacgaatgg 1477ctccatttaa ctttttcttt ttgttcggca g a tac ctt gga gga gca ttt gat 1530gattattctc tccaactgat tcctcttctg tctggaatcc actgccttca gctcttcgtt 1297ttgttgcagt cgttgttgga tgcttttagt agtggaaatg tgtgcgtttc agggatattt 1357gatcacatgc aacattttca ctcataactg gctgaaaaag ttttgcatta atagagctga 1417aatgtctaga tggataagca attgcagtgg tattttaagt acaacatgtg caacgaatgg 1477ctccatttaa ctttttcttt g ttgttcggca the tac ctt gga gga gca ttt gat 1530

Tyr Leu Gly Gly Ala Phe Asp255Tyr Leu Gly Gly Wing Phe Asp255

gtg gag tca cta gta gaa aat ttg ttt aga cag tta gct ggt aac cag 1578val Glu Ser Leu Val Glu Asn Leu Phe Arg Gln Leu Ala Gly Asn Gln260 265 270gtg gag tca cta gta gaa aat ttg ttt aga cag tta gct ggt aac cag 1578val Glu Ser Leu Val Glu Asn Leu Phe Arg Gln Leu Ala Gly Asn Gln260 265 270

gaa ttg gtg gtc aat gtt tat gat gtc aca aac cac tca aac cct ctt 1626Glu Leu Val Val Asn vai Tyr Asp Val Thr Asn His Ser Asn Pro Leu275 280 285gaa ttg gtg gtc aat gtt tat gat gtc aca aac cac tca aac cct ctt 1626Glu Leu Val Val Asn Goes Tyr Asp Val Thr Asn His Ser Asn Pro Leu275 280 285

gtg atg tat gga tct gag gtt cct ctt ggt att ccc tca cca tca cac 1674Val Met Tyr Gly ser Glu Val Pro Leu Gly Ile Pro Ser Pro Ser His290 295 300 305gtg atg tat gga tct gag gtt cct ctt ggt att ccc tca cca tca cac 1674Val Met Tyr Gly be Glu Val Pro Leu Gly Ile Pro Ser Be His290 295 300 305

acc tat acg ttg gat ttt ggt gat cca ttg aga aag cat cag atg gtt 1722Thr Tyr Thr Leu Asp Phe Gly Asp Pro Leu Arg Lys His Gln Met vai310 315 320acc tat acg ttg gat ttt ggt gat cca ttg aga aag cat cag atg gtt 1722Thr Tyr Thr Read Asp Phe Gly Asp Pro Read Arg Lys His Gln Met vai310 315 320

tgc ag gtaaatttgt gtgaattgat cgttggtttt cccattttat attatagaac 1777Cys Argtgc ag gtaaatttgt gtgaattgat cgttggtttt cccattttat attatagaac 1777Cys Arg

gatcggtttt tttaacatcc attggccata aatctgagca g a tac aga aac aag 1831Tyr Arg Asn Lys325gatcggtttt tttaacatcc attggccata aatctgagca g a tac tac aac aag 1831Tyr Arg Asn Lys325

ctt cat gtt tca tgg tct gca att act aca cca tca ggg gtc ttt gtt 1879Leu His vai Ser Trp ser Ala Ile Thr Thr Pro ser Gly Val Phe vai330 335 340ctt cat gtt tca tgg tct gca att act aca cca tca ggg gtc ttt gtt 1879Leu His Will Be Trp Be Wing Ile Thr Thr Pro Be Gly Val Phe vai330 335 340

ata tgt atg ctg gtg ggc tac ata ata tat gct gct tgg agt cgc tac 1927Ile Cys Met Leu vai Gly Tyr Ile Ile Tyr Ala Ala Trp ser Arg Tyr345 350 355gat aat gtt aag gaa gat tgc cgg aaa atg gaa gcg ctg aaa aaa cgg 1975Asp Asn Val Lys Glu Asp Cys Arg Lys Met Glu Ala Leu Lys Lys Arg360 365 370 375ata tgt atg ctg gtg ggc tac ata ata tat gct gct tgg agt cgc tac 1927Ile Cys Met Leu will Gly Tyr Ile Ile Tyr Ala Trp be Arg Tyr345 350 355gat aat gtt aag gaa gat tgc aaa aaa cga aaa Asn Val Lys Glu Asp Cys Lys Arg Lys Met Glu Wing Read Lys Lys Arg360 365 370 375

gca gaa gcg gct gat att gct aaa tct cag gtatagttgg atgttgtttg 2025gca gaa gcg gct gat att gct aaa tct cag gtatagttgg atgttgtttg 2025

Ala Glu Ala Ala Asp Ile Ala Lys ser GlnGlu Wing Wing Asp Wing Ile Wing Lys Be Gln

380 385380 385

cttctctatt tctattgcaa gcttattgtt atatctaaaa ggttcttatt catttatgac 2085ag ttc ctt gca act gtt tct cat gag ate aga aca ccc atg aat ggc 2132Phe Leu Ala Thr Val Ser His Glu Ile Arg Thr Pro Met Asn Gly390 395 400cttctctatt tctattgcaa gcttattgtt atatctaaaa ggttcttatt catttatgac 2085ag ttc ctt gca act gtt tct cat gag till aga aca ccc atg aat ggc 2132Phe Leu Ala Thr Val Be His Glu Ile Arg Thr Pro 39 Asn Gly390

gtg ctg g gtattttctt tgatcttaca acacattcag tttaatgtta tgcaactcat 2189Val Leugtg ctg g gtattttctt tgatcttaca acacattcag tttaatgtta tgcaactcat 2189Val Leu

ttcttttgaa aaaatggaat catctctttg tttcttttcc ctag ga atg ctt gat 2244Gly Met Leu Aspttcttttgaa aaaatggaat catctctttg tttcttttcc ctag ga atg ctt gat 2244Gly Met Leu Asp

405405

atg tta tta gac aca gag ctg aag tca acc cag agg gat tat gca caa 2292Met Leu Leu Asp Thr Glu Leu Lys Ser Thr Gln Arg Asp Tyr Ala Gln410 415 420atg tta tta gac aca gag ctg aag tca acc cag agg gat tat gca caa 2292Met Leu Leu Asp Thr Glu Leu Lys Ser Thr Gln Arg Asp Tyr Ala Gln410 415 420

aca gcc caa gtc tgt gga aag gca tta ata tcc ctg att aac gaa gtg 2340Thr Ala Gln vai Cys Gly Lys Ala Leu Ile Ser Leu Ile Asn Glu Valaca gcc caa gtc tgt gga aag gca tta ata tcc ctg att aac gaa gtg 2340Thr Wing Gln goes Cys Gly Lys Wing Leu Ile Ser Leu Ile Asn Glu Val

425 430 435425 430 435

ctt gac agg gcc aaa ate gag gct ggc aag ata gat ctc gag tca gta 2388Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys Ile Asp Leu Glu Ser Val440 445 450ctt gac agg gcc aaa till gag gct ggc aag ata gat ctc gag tca gta 2388Leu Asp Arg Wing Lys Ile Glu Wing Gly Lys Ile Asp Leu Glu Ser Val440 445 450

cca ttt gac ctg agg tcc ate ctt gat gat gtc ate tcg tta ttt tct 2436Pro Phe Asp Leu Arg Ser Ile Leu Asp Asp vai Ile Ser Leu Phe Ser455 460 465 470cca ttt gac ctg agg tcc until ctt gat gat gtc until tcg tta ttt tct 2436Pro Phe Asp Leu Arg Ser Ile Leu Asp Asp Will Ile Ser Leu Phe Ser455 460 465 470

tca aaa tca aga gag aaa gga att gag gttagttaaa ctgatttcgg 2483tca aaa tca aga gag aaa gga att gag gttagttaaa ctgatttcgg 2483

Ser Lys Ser Arg Glu Lys Gly Ile GluBe Lys Be Arg Glu Lys Gly Ile Glu

475475

tcatggttgg acaaagatca ctaaacgtat taagtttctg ccagccatca attatttctt 2543ttaggaaaat atcatgcact agttccaccg acatctttta gtctcttagc ttgatactct 2603ttccatgaac ttctctgcat taccgtcatg caccatgcac gtttaacttt gtttaatccc 2663agttgatttt cttctatgtt gtaacttcca g ctt gct gta tat gtt tct gaa 2715tcatggttgg acaaagatca ctaaacgtat taagtttctg ccagccatca attatttctt 2543ttaggaaaat atcatgcact agttccaccg acatctttta gtctcttagc ttgatactct 2603ttccatgaac ttctctgcat taccgtcatg caccatgcac gtttaacttt gtttaatccc 2663agttgatttt cttctatgtt gtaacttcca ctt gct gta g gtt tat tct gaa 2715

Leu Ala Val Tyr Val Ser Glu 480 485Leu Wing Val Tyr Val Ser Glu 480 485

aga gtt cct gaa ate ctg ttg ggc gac cct gga agg ttt cgt cag ata 2763Arg Val Pro Glu Ile Leu Leu Gly Asp Pro Gly Arg Phe Arg Gln Ileaga gtt cct gaa till ctg ttg ggc gac cct gga agg ttt cgt cag ata 2763Arg Val Pro Glu Ile Leu Leu Gly Asp Pro Gly Arg Phe Arg Gln Ile

490 495 500490 495 500

att aca aac ttg gtg gga aac tcg ate aag gtaaatgcgc ataacctttg 2813Ile Thr Asn Leu vai Gly Asn ser Ile Lys505 510att aca aac ttg gtg gga aac tcg till aag gtaaatgcgc ataacctttg 2813Ile Thr Asn Leu goes Gly Asn to be Ile Lys505 510

tatccattca tgattttctt taacgatacc aatagttctc accaatgaca tcaggcaact 2873tgtttcttag tatactattg ttcaatgtga acacaag ata aca ata ttt acc ttg 2928Ile Thr Ile Phe Thr Leu15 515tatccattca tgattttctt taacgatacc aatagttctc accaatgaca tcaggcaact 2873tgtttcttag tatactattg ttcaatgtga acacaag ata aca ata ttt acc ttg 2928Ile Thr Ile Phe Thr Leu15 515

tcg cag ttc aca gaa cgg ggg cac att ttt gta caa gtt cac ctg gea 2976Ser Gln Phe Thr Glu Arg Gly His Ile Phe Val Gln vai His Leu Alatcg cag ttc aca gaa cgg ggg cac att ttt gta caa gtt cac ctg gea 2976Ser Gln Phe Thr Glu Arg Gly His Ile Phe Val Gln Goes His Leu Ala

520 525 530520 525 530

gat cac tca aat ctt gea aca gaa gea aaa att gaa cca gta gtc aat 3024Asp His Ser Asn Leu Ala Thr Glu Ala Lys Ile Glu Pro Val Val Asn535 540 545 550gat cac tca aat ctt gea aca gaa gea aaa att gaa cca gta gtc aat 3024Asp His Ser Asn Leu Ala Thr Glu Ala Lys Ile Glu Pro Val Val Asn535 540 545 550

ggg atg aat gga cat aaa gac gag gct att gct ata ccc acc agt ggg 3072Gly Met Asn Gly His Lys Asp Glu Ala Ile Ala Ile Pro Thr Ser Gly555 560 565ggg atg aat gga cat aaa gac gag gct att gct ata ccc acc agg ggg 3072Gly Met Asn Gly His Lys Asp Glu Ala Ile Ala Ile Pro Thr Gly555 560 565

tct cat aac act tta agt ggt ttt gaa gea gct gat age cga aat aac 3120Ser His Asn Thr Leu Ser Gly Phe Glu Ala Ala Asp Ser Arg Asn Asntct cat aac act tta agt ggt ttt gaa gea gct gat age cga aat aac 3120Ser His Asn Thr Leu Ser Gly Phe Glu Wing Asp Ser Arg Asn Asn

570 575 580570 575 580

tgg gaa aac ttc aag ctt ttg ctc tct tac gag aaa aat gaa atg cca 3168Trp Glu Asn Phe Lys Leu Leu Leu Ser Tyr Glu Lys Asn Glu Met Pro 585 590 595tgg gaa aac ttc aag ctt ttg ctc tct tac gag aaa aat gaa atg cca 3168Trp Glu Asn Phe Lys Leu Read Leu Ser Tyr Glu Lys Asn Glu Met Pro 585 590 595

tat gaa agt gat tct gat aaa gta act ctt gtt gtt agt gtg gaa gat 3216Tyr Glu Ser Asp Ser Asp Lys vai Thr Leu Val Val Ser Val Glu Asp600 605 610tat gaa agt gat tct gat aaa gta act ctt gtt gtt agt gtg gaa gat 3216Tyr Glu Ser Asp Ser Asp Lys Go Thr Leu Val Val Ser Val Glu Asp600 605 610

act ggg ata ggc ata cca ctg cat gcc caa ggc cgg gtc ttc acg cct 3264Thr Gly lie Gly lie Pro Leu His Ala Gln Gly Arg Val Phe Thr Pro615 620 625 630act ggg ata ggc ata cca ctg cat gcc caa ggc cgg gtc ttc acg cct 3264Thr Gly lie Gly lie Pro Read his Ala Gln Gly Arg Val Phe Thr Pro615 620 625 630

ttc atg caa gct gac age tca act tet agg aac tat ggt gga act ggc 3312Phe Met Gln Ala Asp Ser Ser Thr Ser Arg Asn Tyr Gly Gly Thr Glyttc atg caa gct gac age tca act tet agg aac tat ggt gga act ggc 3312Phe Met Gln Ala Asp Ser Ser Thr Ser Arg Asn Tyr Gly Gly Thr Gly

635 640 645635 640 645

att gga ttg age ate age aaa tgt ctt gtt gaa ata atg ggt ggt cag 3360Ile Gly Leu Ser Ile ser Lys Cys Leu vai Glu Ile Met Gly Gly Gln 650 655 660att gga ttg age till age aaa tgt ctt gtt gaa ata atg ggt ggt cag 3360Ile Gly Leu Being Ile Being Lys Cys Leu Going Glu Ile Met Gly Gly Gln 650 655 660

ata aac ttt gtc age cga cct ctt gtt ggg age aca ttc aca ttc act 3408Ile Asn Phe vai Ser Arg Pro Leu Val Gly Ser Thr Phe Thr Phe Thraac ttt gtc age cga cct ctt gtt ggg age aca ttc aca ttc act 3408Ile Asn Phe Will Be Arg Pro Read Val Gly Be Thr Phe Thr Phe Thr

665 670 675665 670 675

gct gtt ctg aga agg tgt gac aaa aat gct att agt gac agt aag act 3456Ala Val Leu Arg Arg Cys Asp Lys Asn Ala Ile Ser Asp Ser Lys Thr680 685 690gct gtt ctg aga agg tgt gac aaa aat gct att agt gac agt aag act 3456Ala Val Leu Arg Arg Cys Asp Lys Asn Ala Ile Ser Asp Ser Lys Thr680 685 690

gtt gct ttg cac cca tta ccg tcc agt ttt aaa ggc tta tet gcg cta 3504Val Ala Leu His Pro Leu Pro Ser Ser Phe Lys Gly Leu Ser Ala Leu695 700 705 710gtt gct ttg cac cca tta ccg tcc agt ttt aaa ggc tta tet gcg cta 3504Val Wing Leu His Pro Leu Pro Ser Phe Lys Gly Leu Leu695 700 705 710

ttg gtt gat aaa aga cct gta aga gea act gtg act aag tat cat ttg 3552Leu vai Asp Lys Arg Pro Val Arg Ala Thr Val Thr Lys Tyr His Leuttg gtt gat aaa aga cct gta aga gea act gtg act aag tat cat ttg 3552Leu goes Asp Lys Arg Pro Val Arg Wing Thr Val Thr Lys Tyr His Leu

715 720 725715 720 725

caa agg ctt gga ate act tet gaa gtt gtt ggt acc att gat ccg aca 3600Gln Arg Leu Gly Ile Thr ser Glu vai vai Gly Thr Ile Asp Pro Thr 730 735 740caa agg ctt gga till act tet gaa gtt gtt ggt acc att gat ccg aca 3600Gln Arg Read Gly Ile Thr be Glu go Gly Thr Ile Asp Pro Thr 730 735 740

ttt ggt gtg ttg tet ggg aga aat ggc agt tet cta acc ag 3641ttt ggt gtg ttg tet ggg aga aat ggc agt tet cta acc ag 3641

Phe Gly Val Leu Ser Gly Arg Asn Gly Ser Ser Leu Thr serPhe Gly Val Leu Be Gly Arg Asn Gly Be Be Leu Thr Be

745 750 755745 750 755

gtacttctat cttctacatt cctttcaaaa aattgaaatc ctggagttaa taggctactt 3701ttctctggaa attagaataa acggagcatg cttgcatact aacttcttat gcaaatatca 3761ctgttctatg tataaatgat tacacgataa tgattgtttt ttggtaagta ttactgggta 3821atacttggcc atcatagttc ttgcctttat tttgtatcac ttcggtatat tgctacttct 3881gcggcagttc tctttacgct accgaatgtg atatatttaa ttgaaaattg attttatttt 3941aatctgtgaa aagaacattt ttttaaggcc ccacaattct caaataacta gtaagctgat 4001tgcagatgat atttgaacta tcttgacacc cagttttttt ttttaatcta actcgttttg 4061ttccaaattt gttgtcag c att ggt aag aag cag cca tgc atg ttg cta ate 4113Ile Gly Lys Lys Gln Pro Cys Met Leu Leu Ilegtacttctat cttctacatt cctttcaaaa aattgaaatc ctggagttaa taggctactt 3701ttctctggaa attagaataa acggagcatg cttgcatact aacttcttat gcaaatatca 3761ctgttctatg tataaatgat tacacgataa tgattgtttt ttggtaagta ttactgggta 3821atacttggcc atcatagttc ttgcctttat tttgtatcac ttcggtatat tgctacttct 3881gcggcagttc tctttacgct accgaatgtg atatatttaa ttgaaaattg attttatttt 3941aatctgtgaa aagaacattt ttttaaggcc ccacaattct caaataacta gtaagctgat 4001tgcagatgat atttgaacta tcttgacacc cagttttttt ttttaatcta actcgttttg 4061ttccaaattt gttgtcag c att ggt aag aag cag cca tgc atg ttg cta until 4113Ile Gly Lys Lys Gln Pro Cys Met Read Leu Ile

760 765760 765

gag agt gat tcc tgg gga cca cag atg gat gtc tcc tta cat gct aga 4161Glu Ser Asp Ser Trp Gly Pro Gln Met Asp vai Ser Leu His Ala Arg770 775 780gag agt gat tcc tgg gga cca cag atg gat gtc tcc tta cat gct aga 4161Glu Ser Asp Ser Trp Gly Pro Gln Met Asp Will Be Read His Ala Arg770 775 780

ctt cag gag atg aaa cag agt gat cgc ata cat gta ttg ccc aag gtt 4209Leu Gln Glu Met Lys Gln Ser Asp Arg Ile His Val Leu Pro Lys Valctt cag gag atg aaa cag agt gat cgc ata cat gta ttg ccc aag gtt 4209Leu Gln Glu Met Lys Gln Ser Asp Arg Ile His Val Leu Pro Lys Val

785 790 795785 790 795

ttc ctt ctt tet gct gea gaa tca gac aaa gta aag aag ata cat gea 4257Phe Leu Leu Ser Ala Ala Glu Ser Asp Lys Val Lys Lys Ile His Ala800 805 810 815ttc ctt ctt tet gct gea gaa tca gac aaa gta aag aag ata cat gea 4257Phe Leu Leu Sera Wing Ala Glu Ser Asp Lys Val Lys Ile His Ala800 805 810 815

gtt gat tet gtg ata cca aag cct ctg aaa gea agt gea ctt gcg gcc 4305vai Asp Ser Val Ile Pro Lys Pro Leu Lys Ala Ser Ala Leu Ala Alagt gat tet gtg ata cca aag cct ctg aaa gea agt gea ctt gcg gcc 4305vai Asp Ser Val Ile Pro Lys Pro Leu Lys Ala Ser Ala Leu Ala Ala

820 825 830820 825 830

tgt ctg ttc caa gea ctt ggt ate aca cag ccg age cat gag aaa cgt 4353Cys Leu Phe Gln Ala Leu Gly Ile Thr Gln Pro Ser His Glu Lys Arg835 840 845tgt ctg ttc caa gea ctt ggt till aca cag ccg age cat gag aaa cgt 4353Cys Leu Phe Gln Ala Leu Gly Ile Thr Gln Pro To Be His Glu Lys Arg835 840 845

gac gat tca ggt tet ctt cat ggg cgt gat ggt tca ggt tet ctt cat 4401Asp Asp Ser Gly Ser Leu His Gly Arg Asp Gly Ser Gly Ser Leu His850 855 860gac gat tca ggt tet ctt cat ggg cgt gat ggt tca ggt tet ctt cat 4401Asp Asp Ser Gly Ser Leu His Gly Arg Asp Gly Ser Gly Ser Leu His850 855 860

ggg ttg ctt ctt ggc aag aac ata ttg gta gtt gat gac aac aag gta 4449Gly Leu Leu Leu Gly Lys Asn Ile Leu Val Val Asp Asp Asn Lys Valggg ttg ctt cg ggc aag aac ata ttg gta gtt gat gac aac aag gta 4449Gly Leu Leu Leu Leu Gly Lys Asn Ile Leu Val Val Asp Asp Asn Lys Val

865 870 875865 870 875

aac ctc aga gtg gcc gct ggt aca ttg aag aaa tat ggg gea aag gtg 4497Asn Leu Arg Val Ala Ala Gly Thr Leu Lys Lys Tyr Gly Ala Lys Val880 885 890 895aac ctc aga gtg gcc gct ggt aca ttg aag aaa tat ggg gea aag gtg 4497Asn Leu Arg Val Wing Ala Gly Thr Leu Lys Lys Tyr Gly Ala Lys Val880 885 890 895

gag tgt gtg gag agt gga aaa gat gct ctt tcc ctt cta caa gtg ccg 4545Glu Cys Val Glu Ser Gly Lys Asp Ala Leu Ser Leu Leu Gln vai Pro900 905 910gag tgt gtg gag agt gga aaa gat gct ctt tcc ctt cta caa gtg ccg 4545Glu Cys Val Glu Ser Gly Lys Asp Ala Leu Ser Leu Leu Gln Go Pro900 905 910

cac aag ttt gat ctg tgt ctc atg gac att cag atg ccg gag atg gat 4593His Lys Phe Asp Leu Cys Leu Met Asp Ile Gln Met Pro Glu Met Asp915 920 925cac aag ttt gat ctg tgt ctc atg gac att cag atg ccg gag atg gat 4593His Lys Phe Asp Leu Cys Leu Met Asp Ile Gln Met Pro Glu Met Asp915 920 925

gg gtaagcttat gtcccgttca agatttattt cttttgaatt tgagcctttt 4645gg gtaagcttat gtcccgttca agatttattt cttttgaatt tgagcctttt 4645

GlyGly

atttcatatg atccaaagcg tgacatgtca tctggataag tgccacctcg ttgactatca 4705atttattcac gcaatgaatc aggctttctg cctctttttg aagaaaaaaa aaatgttatt 4765cacacttcgt ggttaatgga aacaagcata tacattgctg acgccaaata ctgattataa 4825ctagtaaaaa tgtcagtctt gtgttagttt tttatgtgcg aaaaatgccg tgtctaattc 4885attatgtgaa aattttcaaa gacattagcc ctgatatcgt cctgcattat gccaaacagt 4945actttaaata ttaatatcag aaggtctaaa atccgtgcag ttaccatttg ttcattgtag 5005ttttaaccat catactgcaa tatgcagttc ttgggctaat gaacataatg gtqtqctaac 5065accctcatga ccaattaact tatgtctctt gaacacttgc tgcatattga catctctgtt 5125cctatatttt ctgaatagta accaagtaat gtcaaaccat gtcattattt tgtcttcgtt 5185taatcaacat agttttgctt catgtgctag a ttt gag gca act cgg caa ata 5237atttcatatg atccaaagcg tgacatgtca tctggataag tgccacctcg ttgactatca 4705atttattcac gcaatgaatc aggctttctg cctctttttg aagaaaaaaa aaatgttatt 4765cacacttcgt ggttaatgga aacaagcata tacattgctg acgccaaata ctgattataa 4825ctagtaaaaa tgtcagtctt gtgttagttt tttatgtgcg aaaaatgccg tgtctaattc 4885attatgtgaa aattttcaaa gacattagcc ctgatatcgt cctgcattat gccaaacagt 4945actttaaata ttaatatcag aaggtctaaa atccgtgcag ttaccatttg ttcattgtag 5005ttttaaccat catactgcaa tatgcagttc ttgggctaat gaacataatg gtqtqctaac 5065accctcatga ccaattaact tatgtctctt gaacacttgc tgcatattga catctctgtt 5125cctatatttt ctgaatagta accaagtaat gtcaaaccat gtcattattt tgtcttcgtt 5185taatcaacat agttttgctt catgtgctag a ttt gag gca act cgg caa ata 5237

Phe Glu Ala Thr Arg Gln Ile930 935Phe Glu Wing Thr Arg Gln Ile930 935

cga gca atg gaa ggg aag gca aat gag cag gca gac gac age gaa tcg 5285Arg Ala Met Glu Gly Lys Ala Asn Glu Gln Ala Asp Asp Ser Glu Sercga gca atg gaa ggg aag gca aat gag cag gca gac gac gac age gaa tcg 5285Arg Ala Met Glu Gly Lys Ala Asn Glu Ala Asp Asp Ser Glu Ser

940 945 950940 945 950

ggt tca gaa ate gca gca aag acg gcc aaa tgg cac ttg cca ate ctg 5333Gly Ser Glu Ile Ala Ala Lys Thr Ala Lys Trp His Leu Pro Ile Leu 955 960 965ggt tca gaa till gca gca aag acg gcc aaa tgg cac ttg cca till ctg 5333Gly Ser Glu Ile Ala Lys Thr Ala Lys Trp His Leu Pro Ile Leu 955 960 965

gca atg acc gct gat gtc ate cag gcc acc cac gag gaa tgc aca aag 5381Ala Met Thr Ala Asp vai Ile Gln Ala Thr His Glu Glu Cys Thr Lysgca atg acc gct gat gtc till cag gcc acc cac gag gaa tgc aca aag 5381Ala Met Thr Wing Asp Go Ile Gln Wing Thr His Glu Glu Cys Thr Lys

970 975 980970 975 980

tgc ggg atg gat ggc tac gtc tcg aag ccc ttt gag gag aag cag ctc 5429Cys Gly Met Asp Gly Tyr Val Ser Lys Pro Phe Glu Glu Lys Gln Leu985 990 995tgc ggg atg gat ggc tac gtc tcg aag ccc ttt gag gag aag cag ctc 5429Cys Gly Met Asp Gly Tyr Val Ser Lys Pro Phe Glu Lys Gln Leu985 990 995

ttc cag gca gta cag aag ttc ttg ggc cca tgc gtt tcc age tga 5474Phe Gln Ala vai Gln 1000 <210> 11<211> 3046<212> DNA<213> Zea mays<220> <221> 5'UTR<222> (l)...(15)<220> <221> CDS<222> (16)..(3003)<220> <221> 3'UTR<222> (3004)..(3046)<400> 11ttc cag gca gta cag aag ttc ttg ggc cca tgc gtt tcc age tga 5474Phe Gln Wing Go Gln 1000 <210> 11 <211> 3046 <212> DNA <213> Zea mays <220> <221> 5'UTR <222> (l) ... (15) <220> <221> CDS <222> (16) .. (3003) <220> <221> 3'UTR <222> (3004) .. (3046) <400> 11

ggggtggagc ctggg atg ggg gtg gga ggc gga ggc gga ggg gag gcc gct 51ggggtggagc ctggg atg ggg gtg gga ggc gga ggc gga ggg gag gcc gct 51

Met Gly vai Gly Gly Gly Gly Gly Gly Glu Ala Ala1 5 10Met Gly goes Gly Gly Gly Gly Gly Gly Gly Glu Wing Wing1 10 10

gcg gtg tcg gcg ccg gcg ccg gcg gag gag gcg ggg aag gac gcg gag 99gcg gtg tcg gcg ccg gcg ccg gcg gag gag gcg ggg aag gac gcg gag 99

Ala vai Ser Ala Pro Ala Pro Ala Glu Glu Ala Gly Lys Asp Ala GluWing Will Be Wing Pro Wing Wing Glu Wing Glu Wing Gly Lys Asp Wing Glu

15 20 2515 20 25

gat ggc ggc ggc tgg acc ttg aag gcg aag ctg ate gcc gta gcg gtg 147gat ggc ggc ggc tgg acc ttg aag gcg aag ctg until gcc gta gcg gtg 147

Asp Gly Gly Gly Trp Thr Leu Lys Ala Lys Leu Ile Ala vai Ala ValAsp Gly Gly Gly Trp Thr Leu Lys Wing Lys Leu Ile Wing Go Wing Val

30 35 4030 35 40

ctg gtg tgg gtg ctg ggg gcc ttg gcg ctc ggg gtg ttc ctg cac tcc 195ctg gtg tgg gtg ctg ggg gcc ttg gcg ctc ggg gtg ttc ctg cac tcc

Leu Val Trp Val Leu Gly Ala Leu Ala Leu Gly Val Phe Leu His SerLeu Val Trp Val Leu Gly Wing Leu Wing Leu Gly Val Phe Leu His Ser

45 50 55 6045 50 55 60

tac ttc cgc cac gcg gcg ctg cgc aag gcg gag gaa ggg ctc gtc age 243tac ttc cgc cac gcg gcg ctg cgc aag gcg gag gaa ggg ctc gtc age 243

Tyr Phe Arg His Ala Ala Leu Arg Lys Ala Glu Glu Gly Leu Val SerTyr Phe Arg His Wing Wing Leu Arg Lys Wing Glu Glu Gly Leu Val Ser

65 70 7565 70 75

atg tgc gag gag cgc gcg cgc atg ctg cag gac cag ttc gcc gtc tcc 291Metatg tgc gag gag cgc gcg cgc atg ctg cag

gtcvaigtcvai

gagGlugagGlu

gccAla125cagGlngccAla125cagGln

15 tggTrp15 tggTrp

tacTyrtacTyr

1010

- 20- 20

2525

ctcLeuctcLeu

agaArg205aatAsnagaArg205aatAsn

30 tctSer30 tctSer

Cys Glu Glu80Cys Glu Glu80

aac cac gtcaac cac gtc

Asn His vai95Asn His vai95

aag cgc ccgaag cgc ccg

Lys Arg Pro110Lys Arg Pro110

agg acg tcgagg acg tcg

Arg Thr serArg thr be

cgg gtg gtgArg Val Valcgg gtg gtgArg Val Val

ate ate aagIle Ile Lys160see you later Ile Lys160

gcg ccg gtcAla Pro Valgcg ccg gtcAla Pro Val

175gac atg atgAsp Met Met190175gac atg atgAsp Met Met190

gea tcg gggAla Ser Glygea tcg gggAla Ser Gly

cac ctg ggtHis Leu Glycac ctg ggtHis Leu Gly

tct gat gccSer Asp Ala240tct gat gccSer Asp Ala240

Arg AlaArg Wing

cac gccHis Alacac gccHis Ala

ccc gcgPro Alaccc gcgPro Wing

ttc gagPhe Glu130cac ggcHis Gly145ttc gagPhe Glu130cac ggcHis Gly145

acc atgThr Metacc atgThr Met

gtt tacVal Tyrgtt tacVal Tyr

tcc ggcSer Glytcc ggcSer Gly

aag geaLys Ala210gtc gtavai vai225aag geaLys Ala210gtc gtavai vai225

aag gagLys Gluaag gagLys Glu

Arg Met Leu Gln85Arg Met Leu Gln85

ctc gcc ate ctcLeu Ala Ile Leu100ctc gcc to ctcLeu Ala Ile Leu100

ctc gac cag aacLeu Asp Gln Asn115ctc gac cag aacLeu Asp Gln Asn115

cgg ccg ctg ctgArg Pro Leu Leucgg ccg ctg ctgArg Pro Leu Leu

Asp Gln Phe Ala Val Ser90Asp Gln Phe Ala Val Ser90

gtc gcc acc ttc cac tacvai Ala Thr Phe His Tyr105gtc gcc acc ttc cac tacvai Wing Thr Phe His Tyr105

gcg gac tac accAla Asp Tyr Thrgcg gac tac accAla Asp Tyr Thr

gac aggAsp Arggac aggAsp Arg

aag cacLys Hisaag cacLys His

tcg cagSer Gln180gag gagGlu Glu195tcg cagSer Gln180gag gagGlu Glu195

gtt cttvai Leugtt cttvai Leu

gag ageGlu Ser150gag ccgGlu Pro165gag ageGlu Ser150gag ccgGlu Pro165

gag accGlu Thrgag accGlu Thr

gac cgtAsp Arggac cgtAsp Arg

act agaThr Argact agaThr Arg

ttg act ttt cctLeu Thr Phe Pro230ttg act ttt cctLeu Thr Phe Pro230

gag gat cgt gttGlu Asp Arg vai245gag gat cgt gttGlu Asp Arg vai245

acg ttcThr Phe120age gggSer Gly135acg ttcThr Phe120age gggSer Gly135

ttc gagPhe Gluttc gagPhe Glu

tct ccgSer Protct ccgSer Pro

gtc tccvai Sergtc tccvai Ser

gag aacGlu Asn200ccg ttcPro Phe215gag aacGlu Asn200ccg ttcPro Phe215

gtc tacvai Tyrgtc tacvai Tyr

gct gccAla Alagct gccAla Wing

gtg gcgvai Alagtg gcgvai Wing

cgc cagArg Glncgc cagArg Gln

gtg cagvai Gln170tac attTyr Ile185gtg cagvai Gln170tac attTyr Ile185

att ttgIle Leuatt ttgIle Leu

tac gcgTyr Ala140cag ggcGln Gly155tac gcgTyr Ala140cag ggcGln Gly155

gat gagAsp Glugat gagAsp Glu

gag gggGlu Glygag gggGlu Gly

agg tcaArg Seragg tcaArg Ser

cgg ctc atg tcgArg Leu Met ser220cgg ctc atg tcgArg Read Met ser220

cat gtc gat cttHis Val Asp Leu235cat gtc gat cttHis Val Asp Leu235

acc gea gga tacThr Ala Gly Tyr250acc gea gga tacThr Wing Gly Tyr250

339339

387387

435435

483483

531531

579579

627627

675675

723723

771ctt ggg gga tca ttt gat gta gaa tca tta gtg gaa aat ttg ttt agg 819Leu Gly Gly Ser Phe Asp vai Glu Ser Leu Val Glu Asn Leu Phe Arg255 260 265771ctt ggg gga tca ttt gat gta gaa tca tta gtg gaa aat ttg ttt agg 819Leu Gly Gly Ser Phe Asp Goes Glu Ser Leu Val Glu Asn Leu Phe Arg255 260 265

cag cta gct ggc aat cag gaa ttg gtg gta aat gtt tat gat gtc aca 867Gln Leu Ala Gly Asn Gln Glu Leu vai vai Asn Val Tyr Asp vai Thr270 275 280cag cta gct ggc aat cag gaa ttg gtg gta aat gtt tat gat gtc aca 867Gln Leu Ala Gly Asn Gln Glu Leu Go Go Asn Val Tyr Asp Go Thr270 275 280

aac agt tcg aac cct ctt gtc atg tat gga tcg gaa gtt tct ctt ggc 915Asn Ser Ser Asn Pro Leu vai Met Tyr Gly Ser Glu Val ser Leu Gly285 290 295 300aac agt tcg aac cct gtc atg tat gga tcg gaa gtt tct ctt ggc 915Asn Ser Ser Asn Pro Leu Goes Met Tyr Gly Ser Glu Val Be Leu Gly285 290 295 300

aac ccc tca cca tcg cac ate tgc atg cta gat ttt ggc gat cca ttc 963Asn Pro Ser Pro Ser His Ile Cys Met Leu Asp Phe Gly Asp Pro Phe305 310 315aac ccc tca cca tcg cac up tgc atg cta gat ttt ggc gat cca ttc 963Asn Pro Ser Pro Ile Cys Met Leu Asp Phe Gly Asp Pro Phe305 310 315

aga aag cat cat atg gtt tgc aga tac aga aac aag cct cag ctc cca 1011Arg Lys His His Met vai Cys Arg Tyr Arg Asn Lys Pro Gln Leu Pro320 325 330aga aag cat cat atg gtt tgc aga tac aga aac aag cct cag ctc cca 1011Arg Lys His His Met Go Cys Arg Tyr Arg Asn Lys Pro Gln Leu Pro320 325 330

tgg tct gea ata tct tcg tca tct ggt gta ttt gtc ata tgt atg ctt 1059Trp Ser Ala Ile Ser ser Ser ser Gly vai Phe Val Ile Cys Met Leu335 340 345tgg tct gea ata tct tcg tca tct ggt gta ttt gtc ata tgt atg ctt 1059Trp Ser Ala Ile Ser Ser Ser Ser Gly Phe Val Ile Cys Met Leu335 340 345

gtg ggg tac ate gtg ggt gcc gct tgg agt cgt tat gat aat gtt aag 1107Val Gly Tyr Ile Val Gly Ala Ala Trp Ser Arg Tyr Asp Asn vai Lys350 355 360gtg ggg tac up gtg ggt gcc gct tgg agt cgt tat gat aat gtt aag 1107Val Gly Tyr Ile Val Gly Wing Trp Ser Arg Tyr Asp Asn goes Lys350 355 360

gaa gat tgc cgg aaa atg gag gag ctg aaa aaa cag gea gaa gea gcc 1155Glu Asp Cys Arg Lys Met Glu Glu Leu Lys Lys Gln Ala Glu Ala Ala365 370 375 380gaa gat tgc cgg aaa atg gag gag ctg aaa aaa cag gea gaa gea gcc 1155Glu Asp Cys Arg Lys Met Glu Glu Leu Lys Lys Gln Ala Glu Ala Ala365 370 375 380

gat gtt gct aaa tct cag ttc ctt gea act gtt tct cat gag ate aga 1203Asp Val Ala Lys ser Gln Phe Leu Ala Thr vai ser His Glu Ile Arg385 390 395gat gtt gct aaa tct cag ttc ctt gea act gtt tct cat gag till aga 1203Asp Val Ala Lys Be Gln Phe Leu Ala Thr Will Be His Glu Ile Arg385 390 395

acg ccc atg aat gga gtt cta ggg atg ctt gat atg ctg tta gac act 1251Thr Pro Met Asn Gly vai Leu Gly Met Leu Asp Met Leu Leu Asp Thr400 405 410acg ccc atg aat gga gtt cta ggg atg ctt gat atg ctg tta gac act 1251Thr Pro Met Asn Gly goes Leu Gly Met Leu Asp Met Leu Asp Thr400 405 410

gac cta acg tcg acc cag agg gat ttt gea caa aca gct caa gtc tgt 1299Asp Leu Thr Ser Thr Gln Arg Asp Phe Ala Gln Thr Ala Gln vai Cys415 420 425gac cta acg tcg acc cag agg gat ttt gea caa aca gct caa gtc tgt 1299Asp Leu Thr Be Thr Gln Arg Asp Phe Ala Gln Thr Ala Gln go Cys415 420 425

gga aag gct tta ata tca cta ate aat gaa gtg ctt gac aga gcg aaa 1347Gly Lys Ala Leu Ile Ser Leu Ile Asn Glu Val Leu Asp Arg Ala Lys430 435 440gga aag gct tta ata tca cta till aat gaa gtg ctt gac aga gcg aaa 1347Gly Lys Ala Leu Ile Ser Leu Ile Asn Glu Val Leu Asp Arg Ala Lys430 435 440

att gaa gcc gga aag ttg gat ctt gag tet gta cca ttt gac ctg aga 1395Ile Glu Ala Gly Lys Leu Asp Leu Glu ser Val Pro Phe Asp Leu Arg445 450 455 460att gaa gcc gga aag ttg gat ctt gag tet gta cca ttt gac ctg aga 1395Ile Glu Ala Gly Lys Leu Asp Leu Glu Be Val Pro Phe Asp Leu Arg445 450 455 460

tcc ate ctt gat gat gtc ate tca tta ttt tet tca aag tca aga gag 1443Ser Ile Leu Asp Asp Val Ile ser Leu Phe Ser Ser Lys ser Arg Glu465 470 475tcc till ctt gat gat gtc till tca tta ttt tet tca aag tca aga gag 1443Ser Ile Leu Asp Asp Val Ile Leu Phe Ser Ser Lys Ser Arg Glu465 470 475

aag gga att gag ctt gct gta tat gtc tet gaa aga gtt cct gaa etc 1491Lys Gly lie Glu Leu Ala Val Tyr Val Ser Glu Arg Val Pro Glu Leuaag gga att gag ctt gct gta tat gtc tet gaa aga gtt cct gaa etc 1491Lys Gly lie Glu Leu Val Wing Tyr Val Ser Glu Arg Val Pro Glu Leu

480 485 490480 485 490

ttg ttg ggt gat cct gga agg ttt cgg cag ata att aca aat tta gtg 1539ttg ttg ggt gat cct gga agg ttt cgg cag ata att aca aat tta gtg 1539

Leu Leu Gly Asp Pro Gly Arg Phe Arg Gln Ile Ile Thr Asn Leu Val495 500 505Leu Leu Gly Asp Pro Gly Arg Phe Arg Gln Ile Ile Thr Asn Leu Val495 500 505

ggc aac tca att aag ttc aca gaa cgg gga cat att ttt gta caa gtt 1587Gly Asn Ser Ile Lys Phe Thr Glu Arg Gly His Ile Phe Val Gln Val510 515 520gg aac tca att aag ttc aca gaa cgg gga cat att ttt gta caa gtt 1587Gly Asn Ser Ile Lys Phe Thr Glu Arg Gly His Ile Phe Val Gln Val510 515 520

cat ctg gea gat cac tca aat ctt gea aca gaa tcc aaa gtt gag tca 1635His Leu Ala Asp His ser Asn Leu Ala Thr Glu Ser Lys Val Glu Ser525 530 535 540cat ctg gea gat cac tca aat ctt gea aca gaa tcc aaa gtt gag tca 1635His Leu Wing Asp His be Asn Leu Wing Ala Thr Glu Ser Lys Val Glu Ser525 530 535 540

gtg gct aac ggg atg aat gga cat aaa gat gag aaa act gct gta gea 1683gtg gct aac ggg atg aat gga cat aaa gat gag aaa act gct gta gea 1683

Val Ala Asn Gly Met Asn Gly His Lys Asp Glu Lys Thr Ala Val Ala545 550 555Val Wing Asn Gly Met Asn Gly His Lys Asp Glu Lys Thr Wing Val Ala545 550 555

acc agt gtt tet etc aac aca cta agt ggt ttt gaa gct gct gat age 1731Thr Ser Val Ser Leu Asn Thr Leu Ser Gly Phe Glu Ala Ala Asp Ser560 565 570acc agt gtt tet etc aac aca cta agt ggt ttt gaa gct gct gat age 1731Thr Ser Val Ser Leu Asn Thr Leu Ser Gly Phe Glu Ala Wing Asp Ser560 565 570

cga aat agt tgg gaa aac ttc aag ctt ttg ctt tet tat gag aaa aat 1779cga aat agt tgg gaa aac ttc aag ctt ttg ctt tet tat gag aaa aat 1779

Arg Asn Ser Trp Glu Asn Phe Lys Leu Leu Leu Ser Tyr Glu Lys Asn575 580 585Arg Asn Be Trp Glu Asn Phe Lys Leu Read Leu Be Tyr Glu Lys Asn575 580 585

gag atg ccc tat gaa agt gta tet gat aaa gtt act ctt gta gta agt 1827Glugag atg ccc tat gta agt gta tet gat aaa gtt act ctt gta gta agt 1827Glu

gtgvai605ttcPhegtgvai605ttcPhe

ggtggt

GlyGly

ggtGlyggtGly

acaThracaThr

agtSer685tctSeragtSer685tctSer

tatTyrtatTyr

gaaGlugaaGlu

ageSerageSer

Met Pro590Met Pro590

gaa gatGlu Aspgaa gatGlu Asp

act cctThr Proact cctThr Pro

act ggaThr Glyact ggaThr Gly

ggt cagGly Gln655ttt actPhe Thr670ggt cagGly Gln655ttt actPhe Thr670

aag cctLys Proaag cctLys Pro

gea ttaAla Leugea ttaAla Leu

cac ttgHis Leucac ttgHis Leu

ttg gctLeu Ala735acg aagThr Lys750ttg gctLeu Ala735acg aagThr Lys750

Tyr Glu Ser Val Ser Asp Lys595Tyr Glu Ser Val Ser Asp Lys595

aca ggg ata ggt ata cca ttgThr Gly Ile Gly Ile Pro Leu610ggg ata ggt ata cca ttgThr Gly Ile Gly Ile Pro Leu610

ttc atgPhe Met625att ggaIle Gly640ttc atgPhe Met625att ggaIle Gly640

ata aaclie Asnok aaclie Asn

gea gctAla Alagea gctAla Wing

gtt atgvai Metgtt atgvai Met

ttg gttLeu vai705caa aggGln Arg720ttg gttLeu vai705caa aggGln Arg720

ctt ggtLeu Glyctt ggtLeu Gly

caa ccaGln Procaa ccaGln Pro

cag gccGln Alacag gccGln Wing

ttg ageLeu Serttg ageLeu Ser

ttt gttPhe Valttt gttPhe Val

ctc caaLeu Gln675ttg cacLeu His690ctc caaLeu Gln675ttg cacLeu His690

gat agaAsp Arggat agaAsp Arg

ctg ggaLeu Glyctg ggaLeu Gly

Qtg ctgvai LeuQtg ctgvai Leu

tgc atgCys Met755tgc atgCys Met755

gat agt tcgAsp Ser Ser630gat agt tcgAsp Ser Ser630

ate age aaaIle Ser Lyseven act aaaIle Ser Lys

645age cga ccaSer Arg Pro660645age cga ccaSer Arg Pro660

aga tgt gacArg Cys Aspaga tgt gacArg Cys Asp

cct ctt ccaPro Leu Procct ctt ccaPro Leu Pro

aga cca gtaArg Pro Val710aga cca gtaArg Pro Val710

att gcc tgcIle Ala Cysatt gcc tgcIle Ala Cys

725tcc ggg agaSer Gly Arg740725tcc ggg agaSer Gly Arg740

tta ttg attLeu Leu lietta ttg attLeu Leu lie

Val Thr Leu600gat gcc caaAsp Ala Gln615Val Thr Leu600gat gcc caaAsp Wing Gln615

act tcc aggThr Ser Argact tcc aggThr Ser Arg

tgt ctt gttCys Leu vaitgt ctt gttCys Leu goes

cat gtt gggHis vai Gly665cat gtt gggHis goes Gly665

aga age gctArg Ser Ala680tcc agt ttcSer Ser Phe695aga age gctArg Ser Ala680tcc agt ttcSer Ser Phe695

aga gct actArg Ala Thraga gct actArg Ala Thr

gat gtt gttAsp vai vaigat gtt gttAsp go go

aat ggc agtAsn Gly Ser745aat ggc agtAsn Gly Ser745

gag agt gatGlu Ser Asp760gag agt gatGlu Ser Asp760

vai Val Serwill Val Ser

gcc aag gtggcc aag gtg

Ala Lys vai620Wing Lys vai620

aca tat ggtaca tat ggt

Thr Tyr GlyThr tyr gly

635635

gaa cta atggaa cta atg

Glu Leu Met650Glu Leu Met650

agt aca ttcagt aca ttc

ser Thr Phebe Thr Phe

att ggt gacIle Gly Aspatt ggt gacIle Gly Asp

aaa ggt ttaLys Gly Leu700aaa ggt ttaLys Gly Leu700

gta act aagvai Thr Lysgta act aagvai Thr Lys

715gct acc attAla Thr Ile730715gct acc attAla Thr Ile730

tct cta accSer Leu Thrtct cta accSer Leu Thr

tca tgg ggcSer Trp Glytca tgg ggcSer Trp Gly

18751875

19231923

19711971

20192019

20672067

21152115

21632163

22112211

22592259

2307ttc aag att gat gta cct tta cga tct cga ctc ctg gag atg aag cag 23552307ttc aag att gat gta cct tta cga tct cga ctc ctg

Phe Lys Il e Asp Val Pro Leu Arg ser Arg Leu Leu Glu Met Lys Gln765 770 775 780Phe Lys Il and Asp Val Pro Read Arg Be Arg Read Leu Read Glu Met Lys Gln765 770 775 780

aat ggt cca cct gga ttg ccc aaa act ate ctt ctc gea gct gea gaa 2403Asn Gly Pro Pro Gly Leu Pro Lys Thr Ile Leu Leu Ala Ala Ala Gluaat ggt cca cct gga ttg ccc aaa act till ctt ctc gea gct gea gaa 2403Asn Gly Pro Pro Gly Leu Pro Lys Thr Ile Leu Leu Ala Ala Ala Glu

785 790 795785 790 795

tcg ggc aaa ctc aaa gea cac tat gea gtt gat tct gtg ate acg aag 2451Ser Gly Lys Leu Lys Ala His Tyr Ala Val Asp Ser vai Ile Thr Lys800 805 810tcg ggc aaa ctc aaa gea cac tat gea gtt gat tct gtg till acg aag 2451Ser Gly Lys Leu Lys Ala His Tyr Ala Val Asp Ser Will Ile Thr Lys800 805 810

cct ctg aaa gea age gga ctt gcc gct tgt cta ttc caa aca ctt ggc 2499Pro Leu Lys Ala Ser Gly Leu Ala Ala Cys Leu Phe Gln Thr Leu Glycct ctg aaa gea age gga ctt gcc gct tgt cta ttc caa aca ctt ggc 2499Pro Leu Lys Ala Ser Gly Leu Cys Wing Leu Phe Gln Thr Leu Gly

815 820 825815 820 825

ate aca cag tca age aac gag aga cgc gac aac tca ggt tcc ctt cat 2547Ile Thr Gln Ser Ser Asn Glu Arg Arg Asp Asn Ser Gly Ser Leu His 830 835 840till aca cag tca acts aac gag aga cgc gac aac tca ggt tcc ctt cat 2547Ile Thr Gln Ser As Asn Glu Arg Arg Asp Asn Ser Gly Ser Leu His 830 835 840

ggg ttg ctc ctt ggc aag aac ata ttg gtg gtt gat gac aac aag gta 2595Gly Leu Leu Leu Gly Lys Asn Ile Leu vai vai Asp Asp Asn Lys vai845 850 855 860ggg ttg ctc ctt ggc aag aac ata ttg gtg gtt gat gac aac aag gta 2595Gly Leu Leu Leu Leu Gly Lys Asn Ile Leu Go Go Asp Asp Asn Lys Go845 850 855 860

aat ctt aga gtg gct gct ggc aca tta aag aaa ttc gga gcg aag gtg 2643 Asn Leu Arg Val Ala Ala Gly Thr Leu Lys Lys Phe Gly Ala Lys Valaat ctt aga gtg gct ggt aca tta aag aaa ttc gga gcg aag gtg 2643 Asn Leu Arg Val Wing Ala Gly Thr Leu Lys Lys Phe Gly Ala Lys Val

865 870 875865 870 875

gag tgc gtg gag agt gga aaa gat gct ctc gcc age cta caa gtt cca 2691Glu Cys Val Glu Ser Gly Lys Asp Ala Leu Ala Ser Leu Gln vai Pro880 885 890gag tgc gtg gag agt gga aaa gat gct ctc gcc age cta caa gtt cca 2691Glu Cys Val Glu Ser Gly Lys Asp Ala Leu Ala Ser Leu Gln Go Pro880 885 890

cat aag ttc cat ctt tgt ctc atg gac att cag atg ccc gaa atg gat 2739His Lys Phe His Leu Cys Leu Met Asp Ile Gln Met Pro Glu Met Aspcat aag ttc cat ct tgt ctc atg gac att cag atg ccc gaa atg gat 2739His Lys Phe His Leu Cys Leu Met Asp Ile Gln Met Pro Glu Met Asp

895 900 905895 900 905

ggg ttc gag gcc acc aag caa ata agg gea atg gaa gcg aag gea aat 2787Gly Phe Glu Ala Thr Lys Gln Ile Arg Ala Met Glu Ala Lys Ala Asn 910 915 920ggg ttc gag gcc acc aag caa ata agg gea atg gaa gcg aag gea aat 2787Gly Phe Glu Ala Thr Lys Gln Ile Arg Ala Met Glu Ala Lys Ala Asn 910 915 920

gag cag gea gtc gcc tgt gac gat tca gat acg gat ggc gcg aca agg 2835Glu Gln Ala Val Ala Cys Asp Asp Ser Asp Thr Asp Gly Ala Thr Arg925 930 935 940gag cag gea gtc gcc tgt gac gat tca gat acg gat ggc gcg aca agg 2835Glu Gln Wing Val Val Wing Cys Asp Asp Ser Asp Thr Asp Gly Ala Thr Arg925 930 935 940

gcg gca aga tgg cac ctg cct gtc ctt gca atg acc gcc gat gtc ate 2883Ala Ala Arg Trp His Leu Pro vai Leu Ala Met Thr Ala Asp vai Ile945 950 955gcg gca aga tgg cac ctg cct gtc ctt gca atg acc gcc gat gtc until 2883Ala Ala Arg Trp His Leu Pro will Leu Ala Met Thr Ala Asp will Ile945 950 955

cag gcc acc cat gag gag tgc aca aag tac ggg atg gat ggg tac gtc 2931Gln Ala Thr His Glu Glu Cys Thr Lys Tyr Gly Met Asp Gly Tyr Valcag gcc acc cat gag gag tgc aca aag tac ggg atg gat ggg tac gtc 2931Gln Wing Thr His Glu Glu Cys Thr Lys Tyr Gly Met Asp Gly Tyr Val

960 965 970960 965 970

acg aag ccc ttc gag gag aag cag ctc ttc cag gcg ctg cag aag ttc 2979Thr Lys Pro Phe Glu Glu Lys Gln Leu Phe Gln Ala Leu Gln Lys Phe 975 980 985acg aag ccc ttc gag gag aag cag ctc ttc cag gcg ctg cag aag ttc 2979Thr Lys Pro Phe Glu Lys Gln Leu Phe Gln Ala Leu Gln Lys Phe 975 980 985

ttg gac cct ggc atg tcc age taa cacccaagtg ctgcgttcgt tgcaagtgag 3033Leu Asp Pro Gly Met Ser Ser990 995ttg gac cct ggc atg tcc age taa cacccaagtg ctgcgttcgt tgcaagtgag 3033Leu Asp Pro Gly Met Ser Ser990 995

gcaccattct cct 3046gcaccattct cct 3046

<210> 12<211> 995<212> PRT<213> Zea mays<400> 12<210> 12 <211> 995 <212> PRT <213> Zea mays <400> 12

Met Gly vai Gly Gly Gly Gly Gly Gly Glu Ala Ala Ala vai Ser Ala15 10 15Met Gly Gly Gly Gly Gly Gly Gly Gly Glu Wing Wing Wing Will Be Wing15 10 15

Pro Ala Pro Ala Glu Glu Ala Gly Lys Asp Ala Glu Asp Gly Gly GlyPro Wing Pro Wing Glu Glu Wing Gly Lys Asp Wing Glu Asp Gly Gly Gly

20 25 3020 25 30

Trp Thr Leu Lys Ala Lys Leu lie Ala Val Ala Val Leu Val Trp vai 35 40 45Trp Thr Leu Lys Wing Lys Leu lie Val Wing Val Wing Val Leu Val Trp goes 35 40 45

Leu Gly Ala Leu Ala Leu Gly vai Phe Leu His Ser Tyr Phe Arg HisLeu Gly Wing Leu Wing Leu Gly will Phe Leu His Be Tyr Phe Arg His

50 55 6050 55 60

Ala Ala Leu Arg Lys Ala Glu Glu Gly Leu vai Ser Met Cys Glu Glu65 70 75 80Wing Wing Leu Arg Lys Wing Glu Glu Gly Leu Will Be Met Cys Glu Glu65 70 75 80

Arg Ala Arg Met Leu Gln Asp Gln Phe Ala Val Ser vai Asn His ValArg Wing Arg Met Leu Gln Asp Gln Phe Wing Val Ser Will Asn His Val

85 90 9585 90 95

His Ala Leu Ala Ile Leu Val Ala Thr Phe His Tyr Glu Lys Arg Pro100 105 110His Wing Leu Wing Ile Leu Val Val Thr Phe His Tyr Glu Lys Arg Pro100 105 110

Pro Ala Leu Asp Gln Asn Thr Phe Ala Asp Tyr Thr Ala Arg Thr SerPro Wing Read Asp Gln Asn Thr Phe Wing Asp Tyr Thr Wing Arg Thr

115 120 125115 120 125

Phe Glu Arg Pro Leu Leu Ser Gly vai Ala Tyr Ala Gln Arg vai vai130 135 140Phe Glu Arg Pro Read Leu Ser Gly Go Wing Tyr Wing Gln Arg Go Go130 135 140

His Gly Asp Arg Glu Ser Phe Glu Arg Gln Gln Gly Trp Ile lie Lys145 150 155 160His Gly Asp Arg Glu Be Phe Glu Arg Gln Gln Gly Trp Ile lie Lys145 150 155 160

Thr Met Lys His Glu Pro Ser Pro Val Gln Asp Glu Tyr Ala Pro Val165 170 175Thr Met Lys His Glu Pro Pro Val Gln Asp Glu Tyr Wing Pro Val165 170 175

vai Tyr Ser Gln Glu Thr vai Ser Tyr Ile Glu Gly Leu Asp Met Met180 185 190will Tyr Be Gln Glu Thr will be Tyr Ile Glu Gly Leu Asp Met Met180 185 190

Ser Gly Glu Glu Asp Arg Glu Asn Ile Leu Arg Ser Arg Ala Ser GlyBe Gly Glu Glu Asp Arg Glu Asn Ile Leu Arg Be Arg Wing Be Gly

195 200 205195 200 205

Lys Ala vai Leu Thr Arg Pro Phe Arg Leu Met Ser Asn His Leu Gly210 215 220Lys Ala Will Read Thr Arg Pro Phe Arg Read Met Ser Asn His Leu Gly210 215 220

Val vai Leu Thr Phe Pro Val Tyr His Val Asp Leu Ser Ser Asp Ala225 230 235 240Val Goes To Read Thr Phe Pro Val Tyr His Val Asp Read To Be Ser Asp Ala225 230 235 240

Lys Glu Glu Asp Arg vai Ala Ala Thr Ala Gly Tyr Leu Gly Gly Ser245 250 255Lys Glu Glu Asp Arg Go Ward Wing Thr Wing Gly Tyr Leu Gly Gly Ser245 250 255

Phe Asp Val Glu Ser Leu Val Glu Asn Leu Phe Arg Gln Leu Ala GlyPhe Asp Val Glu To Be Read Val Glu Asn To Read Phe Arg Gln To Read Wing Gly

260 265 270260 265 270

Asn Gln Glu Leu Val Val Asn Val Tyr Asp Val Thr Asn Ser Ser AsnAsn Gln Glu Leu Val Val Asn Val Tyr Asp Val Thr Asn Ser As Asn

275 280 285275 280 285

Pro Leu vai Met Tyr Gly Ser Glu vai Ser Leu Gly Asn Pro Ser Pro290 295 300Pro Leu Goes Met Tyr Gly Being Glu Will Be Leu Gly Asn Pro Being Pro290 295 300

Ser His Ile Cys Met Leu Asp Phe Gly Asp Pro Phe Arg Lys His His305 310 315 320Be His Ile Cys Met Read Asp Phe Gly Asp Pro Phe Arg Lys His His305 310 315 320

Met vai Cys Arg Tyr Arg Asn Lys Pro Gln Leu Pro Trp Ser Ala Ile325 330 335Met will Cys Arg Tyr Arg Asn Lys Pro Gln Leu Pro Trp Be Wing Ile325 330 335

Ser Ser Ser Ser Gly vai Phe Val Ile Cys Met Leu Val Gly Tyr Ile340 345 350Ser Ser Ser Ser Gly Phe Val Ile Cys Met Leu Val Gly Tyr Ile340 345 350

Val Gly Ala Ala Trp Ser Arg Tyr Asp Asn Val Lys Glu Asp Cys Arg355 360 365Val Gly Wing Trp Wing Be Arg Tyr Asp Asn Val Lys Glu Asp Cys Arg355 360 365

Lys Met Glu Glu Leu Lys Lys Gln Ala Glu Ala Ala Asp Val Ala LysLys Met Glu Glu Leu Lys Lys Gln Wing Glu Wing Wing Asp Val Wing Wing Lys

370 375 380370 375 380

Ser Gln Phe Leu Ala Thr Val Ser His Glu Ile Arg Thr Pro Met Asn385 390 395 400Be Gln Phe Leu Thr Wing Val Be His Glu Ile Arg Thr Pro Met Asn385 390 395 400

Gly Val Leu Gly Met Leu Asp Met Leu Leu Asp Thr Asp Leu Thr serGly Val Leu Gly Met Leu Asp Met Leu Read Asp Thr Asp Leu Thr Be

405 410 415405 410 415

Thr Gln Arg Asp Phe Ala Gln Thr Ala Gln Val Cys Gly Lys Ala Leu420 425 430Thr Gln Arg Asp Phe Wing Gln Thr Wing Gln Val Cys Gly Lys Wing Leu420 425 430

Ile Ser Leu Ile Asn Glu vai Leu Asp Arg Ala Lys Ile Glu Ala Gly435 440 445Ile Ser Leu Ile Asn Glu will Read Asp Arg Wing Lys Ile Glu Wing Gly435 440 445

Lys Leu Asp Leu Glu Ser Val Pro Phe Asp Leu Arg Ser Ile Leu AspLys Leu Asp Leu Glu Be Val Pro Phe Asp Leu Arg Be Ile Leu Asp

450 455 460450 455 460

Asp Val Ile Ser Leu Phe Ser Ser Lys Ser Arg Glu Lys Gly Ile Glu465 470 475 480Asp Val Ile Be Leu Phe Be Be Lys Be Arg Glu Lys Gly Ile Glu465 470 475 480

Leu Ala vai Tyr Val Ser Glu Arg Val Pro Glu Leu Leu Leu Gly AspLeu Ala will Tyr Val Be Glu Arg Val Pro Glu Leu Leu Leu Gly Asp

485 490 495485 490 495

Pro Gly Arg Phe Arg Gln Ile Ile Thr Asn Leu Val Gly Asn Ser Ile500 505 510Pro Gly Arg Phe Arg Gln Ile Ile Thr Asn Read Val Gly Asn Ser Ile500 505 510

Lys Phe Thr Glu Arg Gly His Ile Phe Val Gln Val His Leu Ala Asp515 520 525Lys Phe Thr Glu Arg Gly His Ile Phe Val Gln Val His Leu Wing Asp515 520 525

His Ser Asn Leu Ala Thr Glu Ser Lys Val Glu ser vai Ala Asn GlyHis Ser Asn Read Wing Ala Thr Glu Ser Lys Val Glu Be Go Ala Asn Gly

530 535 540530 535 540

Met Asn Gly His Lys Asp Glu Lys Thr Ala vai Ala Thr ser Val Ser545 550 555 560Met Asn Gly His Lys Asp Glu Lys Thr Wing Go Wing Thr Be Val Ser545 550 555 560

Leu Asn Thr Leu Ser Gly Phe Glu Ala Ala Asp Ser Arg Asn Ser TrpRead Asn Thr Read Le Be Gly Phe Glu Wing Ala Asp Be Arg Asn Be Trp

565 570 575565 570 575

Glu Asn Phe Lys Leu Leu Leu Ser Tyr Glu Lys Asn Glu Met Pro Tyr580 585 590Glu Asn Phe Lys Leu Leu Read Ser Tyr Glu Lys Asn Glu Met Pro Tyr580 585 590

Glu Ser Val Ser Asp Lys Val Thr Leu Val Val Ser Val Glu Asp Thr595 600 605Glu Ser Val Ser Asp Lys Val Thr Leu Val Val Ser Val Glu Asp Thr595 600 605

Gly Ile Gly Ile Pro Leu Asp Ala Gln Ala Lys vai Phe Thr Pro Phe610 615 620Gly Ile Gly Ile Pro Read Asp Wing Gln Wing Lys Go Phe Thr Pro Phe610 615 620

Met Gln Ala Asp ser Ser Thr Ser Arg Thr Tyr Gly Gly Thr Gly Ile625 630 635 640Met Gln Wing Asp Be Be Thr Be Arg Be Tyr Gly Gly Thr Gly Ile625 630 635 640

Gly Leu Ser Ile Ser Lys Cys Leu Val Glu Leu Met Gly Gly Gln Ile5 645 650 655Gly Leu Be Ile Ser Lys Cys Leu Val Glu Leu Met Gly Gly Gln Ile5 645 650 655

Asn Phe Val Ser Arg Pro His Val Gly Ser Thr Phe Thr Phe Thr AlaAsn Phe Val Be Arg Pro His Val Gly Be Thr Phe Thr Phe Thr Wing

660 665 670660 665 670

Ala Leu Gln Arg Cys Asp Arg ser Ala Ile Gly Asp Ser Lys Pro vai675 680 685Wing Read Gln Arg Cys Asp Arg Be Wing Ile Gly Asp Be Lys Pro vai675 680 685

10 Met Leu His Pro Leu Pro Ser Ser Phe Lys Gly Leu Ser Ala Leu Leu690 695 70010 Met Leu His Pro Leu Pro To Be Phe Lys Gly Leu To Be Wing Leu690 695 700

Val Asp Arg Arg Pro vai Arg Ala Thr Val Thr Lys Tyr His Leu Gln705 710 715 720Val Asp Arg Arg Pro Goes Arg Wing Thr Val Thr Lys Tyr His Leu Gln705 710 715 720

Arg Leu Gly Ile Ala Cys Asp vai vai Ala Thr Ile Glu Leu Ala Leu15 725 730 735Arg Leu Gly Ile Wing Cys Asp Goes Go Wing Thr Ile Glu Leu Wing Leu15 725 730 735

Gly Val Leu Ser Gly Arg Asn Gly Ser Ser Leu Thr Ser Thr Lys GlnGly Val Read Be Gly Arg Asn Gly Be Read Le Thr Be Thr Lys Gln

740 745 750740 745 750

Pro Cys Met Leu Leu Ile Glu Ser Asp Ser Trp Gly Phe Lys Ile Asp755 760 765Pro Cys Met Read Leu Ile Glu Be Asp Be Trp Gly Phe Lys Ile Asp755 760 765

k'20 vai Pro Leu Arg Ser Arg Leu Leu Glu Met Lys Gln Asn Gly Pro Pro770 775 780k'20 goes Pro Read Arg Be Arg Read Leu Glu Met Lys Gln Asn Gly Pro Pro770 775 780

Gly Leu Pro Lys Thr Ile Leu Leu Ala Ala Ala Glu Ser Gly Lys Leu785 790 795 800Gly Leu Pro Lys Thr Ile Leu Leu Wing Wing Wing Glu Ser Wing Gly Lys Leu785 790 795 800

Lys Ala His Tyr Ala Val Asp Ser Val Ile Thr Lys Pro Leu Lys Ala25 805 810 815Lys Wing His Tyr Wing Val Asp Ser Val Ile Thr Lys Pro Read Lys Ala25 805 810 815

Ser Gly Leu Ala Ala Cys Leu Phe Gln Thr Leu Gly Ile Thr Gln serSer Gly Leu Wing Cys Wing Phe Phe Gln Thr Read Gly Ile Thr Gln Be

820 825 830820 825 830

Ser Asn Glu Arg Arg Asp Asn ser Gly ser Leu His Gly Leu Leu Leu835 840 845Be Asn Glu Arg Arg Asp Asn Be Gly Be Leu His Gly Leu Leu Leu835 840 845

30 Gly Lys Asn Ile Leu vai vai Asp Asp Asn Lys vai Asn Leu Arg vai850 855 86030 Gly Lys Asn Ile Leu Goes Go Asp Asp Asn Lys Goes Asn Leu Arg Goes 850 855 860

Ala Ala Gly Thr Leu Lys Lys Phe Gly Ala Lys Val Glu Cys Val Glu865 870 875 880Wing Gly Thr Wing Read Lys Lys Phe Gly Wing Lys Val Glu Cys Val Glu865 870 875 880

Ser Gly Lys Asp Ala Leu Ala Ser Leu Gln Val Pro His Lys Phe HisBe Gly Lys Asp Wing Read Wing Wing Be Read Gln Val Pro His Lys Phe His

885 890 895885 890 895

Leu Cys Leu Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala 900 905 910Leu Cys Leu Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Wing 900 905 910

Thr Lys Gln Ile Arg Ala Met Glu Ala Lys Ala Asn Glu Gln Ala vaiThr Lys Gln Ile Arg Wing Met Glu Wing Lys Wing Asn Glu Gln Wing Go

915 920 925915 920 925

Ala Cys Asp Asp Ser Asp Thr Asp Gly Ala Thr Arg Ala Ala Arg Trp930 935 940Cys Wing Asp Asp Be Asp Thr Asp Gly Wing Thr Arg Wing Wing Arg Trp930 935 940

10 His Leu Pro Val Leu Ala Met Thr Ala Asp Val Ile Gln Ala Thr His945 950 955 96010 His Leu Pro Val Leu Wing Met Thr Wing Asp Val Ile Gln Thr Wing His945 950 955 960

Glu Glu Cys Thr Lys Tyr Gly Met Asp Gly Tyr vai Thr Lys Pro PheGlu Glu Cys Thr Lys Tyr Gly Met Asp Gly Tyr Goes Thr Lys Pro Phe

965 970 975965 970 975

Glu Glu Lys Gln Leu Phe Gln Ala Leu Gln Lys Phe Leu Asp Pro Gly 980 985 990Glu Glu Lys Gln Leu Phe Gln Wing Leu Gln Lys Phe Leu Asp Pro Gly 980 985 990

Met Ser Ser995<210> 13<211> 3318 <212> DNAMet Ser Ser995 <210> 13 <211> 3318 <212> DNA

<213> Cucurbita maxima<220><213> Cucurbita maxima <220>

<221> CDS<222> (1)..(2946)25 <220><221> CDS <222> (1) .. (2946) 25 <220>

<221> 3'UTR<222> (2947)..(3318)<400> 13<221> 3'UTR <222> (2947) .. (3318) <400> 13

atg cag gtg age gat aac tet gtg ggt ttg aag tgg aat gag caa atg 48atg cag gtg age gat aac tet gtg ggt ttg aag tgg aat gag caa atg 48

Met Gln vai Ser Asp Asn Ser vai Gly Leu Lys Trp Asn Glu Gln Met1 5 10 15Met Gln Will Be Asp Asn Be Will Gly Leu Lys Trp Asn Glu Gln Met1 5 10 15

gga aca aca aag aag ggt tac aca ttt gtt caa gct aac agg gct tgg 96Gly Thr Thr Lys20gga aca aca aag aag ggt tac aca ttt gtt caa gct aac agg gct tgg 96Gly Thr Thr

aag tatLys Tyr35aag tatLys Tyr35

ate tatIle Tyrsee you Tyr

ctt agaLeu Argctt agaLeu Arg

atg ttaMet Leu50gaa gtgGlu vai65atg ttaMet Leu50gaa gtgGlu vai65

caa ttcGln Phecaa ttcGln Phe

tcc accSer Thrtcc accSer Thr

ttt geaPhe Alattt geaPhe Wing

ggg gtgGly Val130gaa aagGlu Lys145ggg gtgGly Val130gaa aagGlu Lys145

ccc gatPro Aspccc gatPro Asp

ttg gggLeu Glyttg gggLeu Gly

aat gttAsn vaiaat gttAsn will

ttt catPhe His100gaa tacGlu Tyr115ttt catPhe His100gaa tacGlu Tyr115

gcg tatAla Tyrgcg tatAla Tyr

caa cacGln Hishunting cacGln His

cga gatArg Aspcga gatArg Asp

tcg tat att gaaSer Tyr Ile Glu180tcg tat att gaaSer Tyr Ile Glu180

Lys Gly Tyr Thr Phe Val Gln25Lys Gly Tyr Thr Phe Val Gln25

ctt ctg ttc tgg att atg gggLeu Leu Phe Trp Ile Met Gly40ctt ctg ttc tgg att atg gggLeu Leu Phe Trp Ile Met Gly40

aat ggc atg gat gct gat ateAsn Gly Met Asp Ala Asp Ile55aat ggc atg gat gct gat ateAsn Gly Met Asp Wing Asp Ile55

agt atg tgtser Met Cys70agt atg tgtser Met Cys70

agt gtt aacSer Val Asn85agt gtt aacSer Val Asn85

tac ttc aaaTyr Phe Lystac ttc aaaTyr Phe Lys

aca gcc agaThr Ala Argaca gcc agaThr Ala Arg

gea caa agaAla Gln Arg135gea caa agaAla Gln Arg135

ggg tgg atgGly Trp Metggg tgg atgGly Trp Met

150gaa tat geaGlu Tyr Ala165150gaa tat geaGlu Tyr Ala165

tcg ttg gatSer Leu Asptcg ttg gatSer Read Asp

gag cagGlu Glngag cagGlu Gln

cat gttHis vaicat gttHis will

aac cctAsn Pro105act gctThr Ala120aac cctAsn Pro105act gctThr Ala120

gtg attvai Ilegtg attvai Ile

ata agaIle ArgAgaIle Arg

cca gtaPro vaicca gtaPro will

atg atgMet Met185atg atgMet Met185

agg geaArg Alaagg geaArg Wing

75cat gccHis Ala9075cat gccHis Ala90

tet gctSer Alatet gctSer Wing

ttt gaaPhe Gluttt gaaPhe Glu

cat tcgHis Sercat tcgHis Ser

aca atgThr Met155ata tttIle Phe170aca atgThr Met155ata tttIle Phe170

tca ggaSer Glytca ggaSer Gly

Ala Asn Arg30Wing Asn Arg30

atg gcg tttMet Ala Phe45atg gcg tttMet Phe45 Wing

aaa gtg aggLys vai Arg60aaa gtg aggLys will Arg60

agg atg ttgArg Met Leuagg atg ttgArg Met Leu

ttg gct gtcLeu Ala Valttg gct gtcLeu Wing Val

att gat cagIle Asp Gln110att gat cagIle Asp Gln110

cgg cct ctaArg Pro Leucgg cct ctaArg Pro Leu

125gag agg gatGlu Arg Asp140125gag agg gatGlu Arg Asp140

gaa aag gaaGlu Lys Glugaa aag gaaGlu Lys Glu

tet caa gaaSer Gln Glutet caa gaaSer Gln Glu

gag gag gacGlu Glu Asp190gag gag gacGlu Glu Asp190

Ala TrpTrp wing

ate ageIle Seruntil ageIle Ser

agg aatArg Asnagg aatArg Asn

cag gatGln Asp80cag gatGln Asp80

ctt gttLeu Val95ctt gttLeu Val95

gaa actGlu ThrgaG actGlu Thr

etc agtLeu Seretc agtLeu Ser

ate ttcIle PheSee you all Phe

cct tcgPro Ser160aca gtcThr vai175cct tcgPro Ser160aca gtcThr vai175

cgg gaaArg Glucgg gaaArg Glu

144144

192192

240240

288288

336336

384384

432432

480480

528528

576aat att ttg agg gct aga gca aca gga aag gct gtc tta aca aga ccc 624Asn Ile Leu Arg Ala Arg Ala Thr Gly Lys Ala vai Leu Thr Arg Pro576aat att ttg agg gct aga gca aca gga aag gct gtc tta aca aga ccc 624Asn Ile Leu Arg Ala Arg Ala Thr Gly Lys Ala Go Leu Thr Arg Pro

195 200 205195 200 205

ttc agg ctg ctg ggt tcc cat cat ctt gga gtt gtt ttg aca ttt cct 672ttc agg ctg ctg ggt tcc cat cat ctt gga gtt gtt ttg aca ttt cct 672

Phe Arg Leu Leu Gly ser His His Leu Gly vai vai Leu Thr Phe Pro210 215 220Phe Arg Leu Read Gly Be His His Leu Gly Goes Go Leu Thr Phe Pro210 215 220

gtt tac aaa ttc aaa ttg cca tcc ata ccg act gaa gaa gaa cgg ata 720Val Tyr Lys Phe Lys Leu Pro Ser Ile Pro Thr Glu Glu Glu Arg Ile225 230 235 240gtt tac aaa ttc aaa ttg cca tcc ata ccg act gaa gaa gaa cgg ata 720Val Tyr Lys Phe Lys Leu Pro Ser Ile Pro Thu Glu Glu Arg Ile225 230 235 240

gaa gca aca gca ggc tac gtt ggc gga gcc ttt gat gtt gag tca ctc 768Glu Ala Thr Ala Gly Tyr Val Gly Gly Ala Phe Asp Val Glu Ser Leuga gca aca gca ggc tac ggc gga gcc ttt gat gtt gag tca ctc 768Glu Wing Thr Wing Wing Gly Tyr Val Gly Wing Gly Phe Asp Val Glu Ser Leu

245 250 255245 250 255

gtg gag aac ttg ttt ggg cag ctt gca ggg aat cag gcc att ttg gta 816vai Glu Asn Leu Phe Gly Gln Leu Ala Gly Asn Gln Ala Ile Leu Val260 265 270gtg gag aac ttg ttt ggg cag ctt gca ggg aat cag gcc att ttg gta 816vai Glu Asn Leu Phe Gly Gln Leu Wing Gly Asn Gln Wing Ile Leu Val260 265 270

aat gta tat gat gtc acg aac tct tct gat ctt ctc gtg atg tat ggt 864Asn Val Tyr Asp Val Thr Asn Ser Ser Asp Leu Leu Val Met Tyr Glyaat gta tat gat gtc acg aac tct tct gat ctt ctc gtg atg tat ggt 864Asn Val Tyr Asp Val Thr Asn Ser Ser Asp Leu Leu Val Met Tyr Gly

275 280 285275 280 285

cat caa tat caa gat ggt gac ttg tcg ctt tca cat gag age age ctt 912cat cat cat cat cat cat cat cat cat cat cat cat cat cat 912

His Gln Tyr Gln Asp Gly Asp Leu Ser Leu Ser His Glu Ser Ser Leu290 295 300His Gln Tyr Gln Asp Gly Asp Read Be Read Read Be His Glu Be Read Leu290 295 300

gat ttc gga gat cca ttc agg aag cat ttg atg att tgt aga tat cag 960Asp Phe Gly Asp Pro Phe Arg Lys His Leu Met Ile Cys Arg Tyr Gln305 310 315 320gat ttc gga gat cca ttc agg aag cat ttg atg att tgt aga tat cag 960Asp Phe Gly Asp Pro Phe Arg Lys His Leu Met Ile Cys Arg Tyr Gln305 310 315 320

cag agg gct ccc aca tcc tgg act gcc cta act act gca ttc tta ttc 1008Gln Arg Ala Pro Thr ser Trp Thr Ala Leu Thr Thr Ala Phe Leu Phecag agg gct ccc aca tcc tgg act gcc cta act act gca ttc tta ttc 1008Gln Arg Ala Pro Thr be Trp Thr Ala Leu Thr Thr Ala Phe Leu Phe

325 330 335325 330 335

ttc gtg ate ggt ttg tta gtt gga tat att ttg tat ggt gca gca act 1056Phe vai Ile Gly Leu Leu Val Gly Tyr Ile Leu Tyr Gly Ala Ala Thr340 345 350ttc gtg till ggt ttg tta gtt gga tat att ttg tat ggt gca gca act 1056Phe goes Ile Gly Leu Leu Val Gly Tyr Ile Leu Tyr Gly Wing Ala Thr340 345 350

cac att gtg aag gtt gaa gat gat ttt cat gaa atg caa gta ctg aaa 1104His Ile Val Lys vai Glu Asp Asp Phe His Glu Met Gln Val Leu Lys355 360 365cac att gtg aag gtt gaa gat gat ttt cat gaa atg caa gta ctg aaa 1104His Ile Val Lys Goes Glu Asp Asp Phe His Glu Met Gln Val Leu Lys355 360 365

gtt cga gcg gag gct gcc gat gta gca aaa tcc cag ttt ctt gca act 1152vai Arg Ala Glu Ala Ala Asp Val Ala Lys Ser Gln Phe Leu Ala Thr370 375 380gtt cga gcg gag gct gcc gat gta gca aaa tcc cag ttt ctt gca act 1152vai Arg Wing Glu Wing Wing Asp Val Wing Lys Ser Gln Phe Leu Wing Thr370 375 380

gtt tct cat gaa att agg aca cca atg aat ggc ate ctc gga atg ctt 1200Val ser His Glu lie Arg Thr Pro Met Asn Gly Ile Leu Gly Met Leu385 390 395 400gtt tct cat gaa att agg aca cca atg aat ggc up to ctc gga atg ctt 1200Val be His Glu lie Arg Thr Pro Met Asn Gly Ile Leu Gly Met Leu385 390 395 400

gct ctg ctt ctg gat aca gat cta agt tcc aca cag aag gat tat gct 1248Ala Leu Leu Leu Asp Thr Asp Leu Ser Ser Thr Gln Lys Asp Tyr Ala405 410 415gct ctg ctt ctg gat aca gat cta agt tcc aca cag aag gat tat gct 1248Ala Leu Leu Leu Asp Thr Asp Leu Being Ser Thr Gln Lys Asp Tyr Ala405 410 415

caa act gcc cag gct tgt gga aag gca ttg ata gca tta ata aat gag 1296Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu Ile Ala Leu Ile Asn Glucaa act gcc cag gct tgt gga aag gca ttg ata gca tta ata aat gag 1296Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu Ile Ala Leu Ile Asn Glu

420 425 430420 425 430

gtt ctt gac cgg gca aaa att gaa gct gga aag tta gaa ctg gaa gca 1344Val Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys Leu Glu Leu Glu Ala435 440 445gtt ctt gac cgg gca aaa att gaa gct gga aag tta gaa ctg gaa gca 1344Val Leu Asp Arg Wing Lys Ile Glu Wing Gly Lys Leu Glu Leu Glu Ala435 440 445

gtt cca ttc gac att cga tca ata ctt gat gac gtg cta tct tta ttt 1392vai Pro Phe Asp Ile Arg Ser Ile Leu Asp Asp Val Leu Ser Leu Phe450 455 460gtt cca ttc gac att cga tca ata ctt gat gac gtg cta tct tta ttt 1392vai Pro Phe Asp Ile Arg Ser Ile Leu Asp Asp Val Leu Ser Leu Phe450 455 460

tcc gag aag tcc aga caa aag ggt ctg gag ctg gca gtt ttt gtt tct 1440Ser Glu Lys Ser Arg Gln Lys Gly Leu Glu Leu Ala vai Phe vai Ser465 470 475 480tcc gag aag tcc aga caa aag ggt ctg gag ctg gca gtt ttt gtt tct 1440Ser Glu Lys Be Arg Gln Lys Gly Leu Glu Leu Wing Go Phe Go Ser465 470 475 480

gat aaa gtt cca gaa att gta att gga gat cct gga aga ttc aga caa 1488Asp Lys Val Pro Glu Ile vai Ile Gly Asp Pro Gly Arg Phe Arg Gln485 490 495gat aaa gtt cca gaa att gta att gga gat cct gga aga ttc aga caa 1488Asp Lys Val Pro Glu Ile Go Ile Gly Asp Pro Gly Arg Phe Arg Gln485 490 495

att ata aca aat ctt gtg ggt aac tct gtt aag ttt act gaa aga gga 1536Ile Ile Thr Asn Leu vai Gly Asn ser Val Lys Phe Thr Glu Arg Glyatt ata aca aat ctt gtg ggt aac tct gtt aag ttt act gaa aga gga 1536Ile Ile Thr Asn Leu Goes Gly Asn Be Val Lys Phe Thr Glu Arg Gly

500 505 510500 505 510

cat ata ttt gtt aaa gta cac cta gct gag aat tca aaa gtc tcc atg 1584His Ile Phe vai Lys Val His Leu Ala Glu Asn ser Lys vai Ser Met515 520 525cat ata ttt gtt aaa gta cac cta gct gag aat tca aaa gtc tcc atg 1584His Ile Phe Will Lys Val His Leu Ala Glu Asn Be Lys Will Be Met515 520 525

gac tcg gaa tac gtc aac gga ata tcc gac agt ggc tta ttc gta ttg 1632Asp Ser Glu Tyr vai Asn Gly Ile ser Asp Ser Gly Leu Phe Val Leugac tcg gaa tac gtc aac gga ata tcc gac agt ggc tta ttc gta ttg 1632Asp Ser Glu Tyr will Asn Gly Ile be Asp Ser Gly Leu Phe Val Leu

530 535 540530 535 540

gat ggt cgt gaa ttt caa act ttg agt gga cgc gag gca gcc gat gat 1680Asp Gly Arg Glu Phe Gln Thr Leu Ser Gly Arg Glu Ala Ala Asp Asp5 545 550 555 560gat ggt cgt gaa ttt caa act ttg agt gga cgc gag gca gcc gat gat 1680Asp Gly Arg Glu Phe Gln Thr Leu Ser Gly Arg Glu Wing Asp Asp5 545 550 555 560

cag aac agt tgg gat aac ttc aag cat cta ate gct gac gac aac ttc 1728Gln Asn Ser Trp Asp Asn Phe Lys His Leu Ile Ala Asp Asp Asn Phecag aac agt tgg gat aac ttc aag cat cta to gct gac gac aac ttc 1728Gln Asn Ser Trp Asp Asn Phe Lys His Leu Ile Ala Asp Asp Asn Phe

565 570 575565 570 575

cag tcg aat gcc gct tca aac aac tca gca gtt acc aac aag ggt tgt 177610 Gln Ser Asn Ala Ala Ser Asn Asn Ser Ala Val Thr Asn Lys Gly Cys580 585 590cag tcg aat gcc gct tca aac aac tac gca gtt acc aac aag ggt tgt 177610 Gln Ser Asn Wing Ala Ser Asn Asn Ser Wing Val Thr Asn Lys Gly Cys580 585 590

gat cat gtt act ttg atg gta agt gtg gag gat act gga att ggg ate 1824Asp His vai Thr Leu Met Val Ser Val Glu Asp Thr Gly Ile Gly Ile595 600 605gat cat gtt act ttg atg gta agt gtg gag gat act gga att ggg until 1824Asp His will Thr Leu Met Val Ser Val Glu Asp Thr Gly Ile Gly Ile595 600 605

15 ctt tta cat gcc caa aat cga gtt ttc aca ccc ttc atg caa gca gat 1872Leu Leu His Ala Gln Asn Arg vai Phe Thr Pro Phe Met Gln Ala Asp15 ctt tta cat gcc caa aat cga gtt ttc aca ccc ttc atg caa gca gat 1872Leu Read His Ala Gln Asn Arg Goes Phe Thr Pro Phe Met Gln Ala Asp

610 615 620610 615 620

age tcg acc tcc cga aat tat gga ggg act ggt att ggt ttg agt ate 1920Ser Ser Thr Ser Arg Asn Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile20 625 630 635 640age tcg acc tcc cga aat tat ggg ggt act ggt att ggt ttg agt until 1920Ser Ser Thr Be Arg Asn Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile20 625 630 635 640

age aaa tgt tta gtt gag tta atg ggt ggt cag ate aac ttc ata age 1968ser Lys Cys Leu Val Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Serage aaa tgt tta gtt gag tta atg ggt ggt cag till aac ttc ata age 1968ser Lys Cys Leu Val Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Ser

645 650 655645 650 655

cgg cct cag att gga age acg ttt tcc ttc act gct gta ttt gga aaa 201625 Arg Pro Gln Ile Gly Ser Thr Phe Ser Phe Thr Ala Val Phe Gly Lys660 665 670cgg cct cag att gga age acg ttt tcc ttc act gct gta ttt gga aaa 201625 Arg Pro Gln Ile Gly Be Thr Phe Be Ala Val Phe Gly Lys660 665 670

tgt aag aaa aac tcg atg aat gat atg aaa aag ccc aac tet gaa gaa 2064Cys Lys Lys Asn Ser Met Asn Asp Met Lys Lys Pro Asn Ser Glu Glu675 680 685tgt aag aaa aac tcg ag gat atg aaa aag ccc aac tet gaa gaa 2064Cys Lys Lys Asn Ser Met Asn Asp Met Lys Lys Pro Asn Ser Glu Glu675 680 685

30 ctt ccc ccc agt ttt aga gga atg aaa gca ata gta gtt gat age aaa 2112Leu Pro Pro Ser Phe Arg Gly Met Lys Ala Ile Val Val Asp Ser Lys690 695 700cat gta cga gct tct gta acc agg tat cat ttg aag aga ctt ggt ate 2160His vai Arg Ala Ser Val Thr Arg Tyr His Leu Lys Arg Leu Gly Ile705 710 715 72030 ctt ccc ccc agt ttt aga gga atg aaa gca ata gta gtt gat age aaa 2112Leu Pro Pro Ser Phe Arg Gly Met Lys Ala Ile Val Val Asp Ser Lys690 695 700cat gta cga gct tct gta acc cat ttg aag aga ctt ggt till 2160His Will Arg Wing Be Val Thr Arg Tyr His Leu Lys Arg Leu Gly Ile705 710 715 720

ata gtt gaa gtc acc aat age ate aac atg gea gct tct tta ttc aga 2208Ile Val Glu Val Thr Asn Ser Ile Asn Met Ala Ala ser Leu Phe Arg725 730 735Ata Gtt Gaa Gtc Acc Aat Age Aac Atg Gea Gct Tct Tta Ttc Aga 2208Ile Val Glu Val Thr Asn Ser Ile Asn Met Wing Ala Leu Phe Arg725 730 735

gaa aat gga tcc aca ctg cca aga aac aca ate ctt cca gat atg ate 2256Glu Asn Gly Ser Thr Leu Pro Arg Asn Thr Ile Leu Pro Asp Met Ile740 745 750gaa aat gga tcc aca ctg cca aga aac aca up to ctt cca gat atg up to 2256Glu Asn Gly Ser Thr Leu Pro Arg Asn Thr Ile Leu Pro Asp Met Ile740 745 750

tta gtt gaa aag gac ata cta aat tct gat gag gaa tgt ggg ate att 2304Leu Val Glu Lys Asp Ile Leu Asn Ser Asp Glu Glu Cys Gly Ile Ile755 760 765tta gtt gaa aag gac ata cta aat tct gat gag gaa tgt ggg until att 2304Leu Val Glu Lys Asp Ile Leu Asn Ser Asp Glu Glu Cys Gly Ile Ile755 760 765

cat cat ctg aac tgg aaa ccg aac ggt agt tcg gtt aag ttt cca aag 2352His His Leu Asn Trp Lys Pro Asn Gly Ser Ser vai Lys Phe Pro Lys770 775 780cat cat ctg aac tgg aaa ccg aac ggt agt tcg gtt aag ttt cca aag 2352His His Leu Asn Trp Lys Pro Asn Gly Ser Ser Will Lys Phe Pro Lys770 775 780

ctg ate ctt ctc gct acc aat att gcc act gct gaa cta gac aag gea 2400Leu Ile Leu Leu Ala Thr Asn Ile Ala Thr Ala Glu Leu Asp Lys Ala785 790 795 800ctg until ctt ctc gct acc aat att gcc act gct gaa cta gac aag gea 2400Leu Ile Leu Leu Thr Wing Asn Ile Ala Thr Wing Glu Leu Asp Lys Ala785 790 795 800

aga gea gea ggt ttt gea gac acc gtg ate atg aag ccg ttg agg gcg 2448Arg Ala Ala Gly Phe Ala Asp Thr Val Ile Met Lys Pro Leu Arg Ala805 810 815aga gea gea ggt ttt gea gac acc gtg up tog aag ccg ttg agg gcg 2448Arg Ala Gly Phe Ala Asp Thr Val Ile Met Lys Pro Leu Arg Ala805 810 815

act atg gtg gct gcc tgt ctt caa caa gta ctc ggg gtt aag aat cag 2496Thr Met Val Ala Ala Cys Leu Gln Gln Val Leu Gly Val Lys Asn Gln820 825 830act atg gtg gct gcc tgt ctt caa caa gta ctc ggg gtt aag aat cag 2496Thr Met Val Wing Cys Wing Leu Gln Val Leu Gly Val Lys Asn Gln820 825 830

aga cgg ccg aat ggt tct gct ttc ctc cag age ctt ctc tgt ggc aag 2544Arg Arg Pro Asn Gly Ser Ala Phe Leu Gln Ser Leu Leu Cys Gly Lys835 840 845agg cgg ccg aat ggt tct gct ttc ctc cag age ctt ctc tgt ggc aag 2544Arg Arg Pro Asn Gly Ser Ala Phe Leu Gln Ser Leu Cys Gly Lys835 840 845

aga ate tta att gtt gat gac aac cga gta aac cgt cgg gtc gct gea 2592Arg Ile Leu Ile vai Asp Asp Asn Arg Val Asn Arg Arg vai Ala Ala850 855 860aga till tta att gtt gat gac aac cga gta aac cgt cgg gtc gct gea 2592Arg Ile Leu Ile Go Asp Asp Asn Arg Val Asn Arg Arg Go Ala Ala850 855 860

ggc gct ctg aag aaa ttt ggt gea gat gtt gag tgt gea gat age ggg 2640Gly Ala Leu Lys Lys Phe Gly Ala Asp Val Glu Cys Ala Asp Ser Gly865 870 875 880ggc gct ctg aag aaa ttt ggt gea gat gtt gag tgt gea gat age ggg 2640Gly Wing Leu Lys Lys Phe Gly Wing Asp Val Glu Cys Wing Asp Ser Gly865 870 875 880

aaa tct gca ctg aág ttg ctt cag cta ccg cat aat ttt gat gct tgc 2688Lys Ser Ala Leu Lys Leu Leu Gln Leu Pro His Asn Phe Asp Ala Cys885 890 895aaa tct gca ctg aag ttg ctt cag cta ccg cat aat ttt gat gct tgc 2688Lys Ser Ala Leu Lys Leu Leu Gln Leu Pro His Asn Phe Asp Ala Cys885 890 895

ttc atg gat att caa atg cct gaa atg gat ggg ttt gag gcg act cgt 27B6Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala Thr Argttc atg gat att caa atg cct gaa atg gat ggg ttt gag gcg act cgt 27B6Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala Thr Arg

900 905 910900 905 910

cgt ate agg aca atg gag gtc gag gca aac aaa gga gga ttg tct gca 2784Arg Ile Arg Thr Met Glu vai Glu Ala Asn Lys Gly Gly Leu Ser Ala 915 920 925cgt till agg aca atg gag gtc gag gca aac aaa gga gga ttg tct gca 2784Arg Ile Arg Thr Met Glu Go Glu Ala Asn Lys Gly Gly Leu Ser Ala 915 920 925

aca gaa ggc aaa cgg cct ata cca ata tta gca atg act gca gac gtg 2832Thr Glu Gly Lys Arg Pro Ile Pro lie Leu Ala Met Thr Ala Asp vaiaca gaa ggc aaa cgg cct ata cca ata tta gca atg act gca gac gtg 2832Thr Glu Gly Lys Arg Pro Ile Pro lie

930 935 940930 935 940

att cat gct acg tac gaa gaa tgc ctg aaa tgc ggt atg aat ggt tac 2880Ile His Ala Thr Tyr Glu Glu Cys Leu Lys Cys Gly Met Asn Gly Tyr945 950 955 960att cat gct acg tac gaa gaa tgc ctg aaa tgc ggt atg aat ggt tac 2880Ile His Wing Thr Tyr Glu Glu Cys Leu Lys Cys Gly Met Asn Gly Tyr945 950 955 960

gtc tcg aaa ccc ttt gaa gaa gaa aat cta tac aag gaa gtt gcc cga 2928Val Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr Lys Glu Val Ala Arg965 970 975gtc tcg aaa ccc ttt gaa gaa gaa aat cta tac aag gaa gtt gcc cga 2928Val Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr Lys Glu Val Wing Arg965 970 975

ttt ttc aaa aaa cca tag tccatcaaaa gcttcatgaa tgacaagagg 2976Phe Phe Lys Lys Pro980ttt ttc aaa aaa cca tag tccatcaaaa gcttcatgaa tgacaagagg 2976Phe Phe Lys Lys Pro980

tcatcagctg tagagctcct tttggtgggt tggaaagtac cagcaagttt tgacaccatt 3036gctggtgcta actgtcctgt tgctggtgac cgaggaaccg agttcaacga gggcgtcggg 3096aattctcgac tgccattgag actcgggttt gagctgctac catttctaac caactaaata 3156ttttattttg actagaatgt gagtacctac ctgtatacta caccagaaat atccatcccc 3216aaatggatgt ataattatgg ttgcaagggg aaggagctaa attgtaaatg ctcatatttc 3276taagacctct tctaaacatc tttatagttg gttggcttag gc 3318tcatcagctg tagagctcct tttggtgggt tggaaagtac cagcaagttt tgacaccatt 3036gctggtgcta actgtcctgt tgctggtgac cgaggaaccg agttcaacga gggcgtcggg 3096aattctcgac tgccattgag actcgggttt gagctgctac catttctaac caactaaata 3156ttttattttg actagaatgt gagtacctac ctgtatacta caccagaaat atccatcccc 3216aaatggatgt ataattatgg ttgcaagggg aaggagctaa attgtaaatg ctcatatttc 3276taagacctct tctaaacatc tttatagttg gttggcttag gc 3318

<210> 14<211> 981<212> PRT<210> 14 <211> 981 <212> PRT

<213> Cucurbita maxima<400> 14<213> Cucurbita maxima <400> 14

Met Gln Val Ser Asp Asn Ser vai Gly Leu Lys Trp Asn Glu Gln Met15 10 15Met Gln Val Ser Asp Asn Ser Go Gly Leu Lys Trp Asn Glu Gln Met15 10 15

Gly Thr Thr Lys Lys Gly Tyr Thr Phe Val Gln Ala Asn Arg Ala Trp5 20 25 30Gly Thr Thr Lys Lys Gly Tyr Thr Phe Val Gln Wing Asn Arg Wing Trp5 20 25 30

Leu Arg Lys Tyr Leu Leu Phe Trp Ile Met Gly Met Ala Phe Ile SerLeu Arg Lys Tyr Leu Read Phe Trp Ile Met Gly Met Wing Phe Ile Ser

35 40 4535 40 45

Met Leu Ile Tyr Asn Gly Met Asp Ala Asp Ile Lys vai Arg Arg Asn50 55 60Met Leu Ile Tyr Asn Gly Met Asp Wing Asp Ile Lys Goes Arg Arg Asn50 55 60

10 Glu Val Leu Gly Ser Met Cys Glu Gln Arg Ala Arg Met Leu Gln Asp65 70 75 8010 Glu Val Leu Gly Ser Met Cys Glu Gln Arg Wing Arg Met Leu Gln Asp65 70 75 80

Gln Phe Asn Val Ser vai Asn His vai His Ala Leu Ala Val Leu ValGln Phe Asn Val Ser Goes Asn His Goes His Wing Leu Wing Val Leu Val

85 90 9585 90 95

Ser Thr Phe His Tyr Phe Lys Asn Pro Ser Ala Ile Asp Gln Glu Thr15 100 105 110Ser Thr Phe His Tyr Phe Lys Asn Pro Ala Ile Asp Gln Glu Thr15 100 105 110

Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe Glu Arg Pro Leu Leu SerPhe Wing Glu Tyr Thr Wing Arg Thr Wing Phe Wing Glu Arg Pro Read Leu Ser

115 120 125115 120 125

Gly Val Ala Tyr Ala Gln Arg vai Ile His Ser Glu Arg Asp Ile Phe130 135 140Gly Val Wing Tyr Wing Gln Arg Goes Ile His Being Glu Arg Asp Ile Phe130 135 140

' 20 Glu Lys Gln His Gly Trp Met Ile Arg Thr Met Glu Lys Glu Pro Ser145 150 155 160'20 Glu Lys Gln His Gly Trp Met Ile Arg Thr Met Glu Lys Glu Pro Ser145 150 155 160

Pro Asp Arg Asp Glu Tyr Ala Pro vai Ile Phe Ser Gln Glu Thr vaiPro Asp Arg Asp Glu Tyr Wing Pro Go Ile Phe Ser Gln Glu Thr Go

165 170 175165 170 175

Ser Tyr Ile Glu Ser Leu Asp Met Met Ser Gly Glu Glu Asp Arg Glu25 180 185 190Be Tyr Ile Glu Be Read Asp Met Met Be Gly Glu Glu Asp Arg Glu25 180 185 190

Asn Ile Leu Arg Ala Arg Ala Thr Gly Lys Ala Val Leu Thr Arg ProAsn Ile Leu Arg Wing Arg Wing Thr Wing Gly Lys Wing Val Leu Thr Arg Pro

195 200 205195 200 205

Phe Arg Leu Leu Gly Ser His His Leu Gly Val Val Leu Thr Phe Pro210 215 220Phe Arg Read Leu Gly Be His His Read Le Gly Val Val Leu Thr Phe Pro210 215 220

30 Val Tyr Lys Phe Lys Leu Pro Ser Ile Pro Thr Glu Glu Glu Arg Ile225 230 235 24030 Val Tyr Lys Phe Lys Leu Pro Ser Ile Pro Thr Thru Glu Glu Arg Ile225 230 235 240

Glu Ala Thr Ala Gly Tyr Val Gly Gly Ala Phe Asp vai Glu Ser Leu245 250 255Glu Wing Thr Wing Gly Tyr Val Gly Gly Wing Phe Asp will Glu Be Leu245 250 255

Val Glu Asn Leu Phe Gly Gln Leu Ala Gly Asn Gln Ala Ile Leu ValVal Glu Asn Leu Phe Gly Gln Leu Wing Gly Asn Gln Wing Ile Leu Val

260 265 270260 265 270

Asn Val Tyr Asp vai Thr Asn ser ser Asp Leu Leu Val Met Tyr Gly275 280 285Asn Val Tyr Asp Will Thr Asn Be Being Asp Leu Leu Val Met Tyr Gly275 280 285

His Gln Tyr Gln Asp Gly Asp Leu ser Leu Ser His Glu Ser Ser LeuHis Gln Tyr Gln Asp Gly Asp Read Be Read Read Be His Glu Be Read Read

290 295 300290 295 300

Asp Phe Gly Asp Pro Phe Arg Lys His Leu Met Ile Cys Arg Tyr Gln305 310 315 320Asp Phe Gly Asp Pro Phe Arg Lys His Leu Met Ile Cys Arg Tyr Gln305 310 315 320

Gln Arg Ala Pro Thr Ser Trp Thr Ala Leu Thr Thr Ala Phe Leu PheGln Arg Ward Pro Thr Be Trp Thr Wing Read Thr Thr Wing Phe Leu Phe

325 330 335325 330 335

Phe Val Ile Gly Leu Leu vai Gly Tyr Ile Leu Tyr Gly Ala Ala ThrPhe Val Ile Gly Leu Leu Go Gly Tyr Ile Leu Tyr Gly Wing Ward Thr

340 345 350340 345 350

His Ile vai Lys Val Glu Asp Asp Phe His Glu Met Gln vai Leu Lys355 360 365His Ile goes Lys Val Glu Asp Asp Phe His Glu Met Gln goes Leu Lys355 360 365

Val Arg Ala Glu Ala Ala Asp vai Ala Lys Ser Gln Phe Leu Ala ThrVal Arg Wing Glu Wing Wing Asp goes Wing Lys Ser Gln Phe Leu Wing Thr

370 375 380370 375 380

Val Ser His Glu Ile Arg Thr Pro Met Asn Gly Ile Leu Gly Met Leu385 390 395 400Val Ser His Glu Ile Arg Thr Pro Met Asn Gly Ile Leu Gly Met Leu385 390 395 400

Ala Leu Leu Leu Asp Thr Asp Leu Ser ser Thr Gln Lys Asp Tyr AlaWing Read Leu Read Asp Thr Asp Read Be Be Thr Gln Lys Asp Tyr Wing

405 410 415405 410 415

Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu Ile Ala Leu Ile Asn GluGln Thr Wing Gln Cys Wing Gly Lys Wing Leu Ile Wing Leu Ile Asn Glu

420 425 430420 425 430

Val Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys Leu Glu Leu Glu Ala435 440 445Val Leu Asp Arg Wing Lys Ile Glu Wing Gly Lys Leu Glu Leu Glu Ala435 440 445

vai Pro Phe Asp Ile Arg ser Ile Leu Asp Asp Val Leu Ser Leu Phego to Phe Asp Ile Arg be Ile Leu Asp Asp Val Leu Be Leu Phe

450 455 460450 455 460

Ser Glu Lys ser Arg Gln Lys Gly Leu Glu Leu Ala Val Phe Val Ser465 470 475 480Be Glu Lys Be Arg Gln Lys Gly Leu Glu Leu Wing Val Phe Val Ser465 470 475 480

Asp Lys vai Pro Glu Ile Val Ile Gly Asp Pro Gly Arg Phe Arg GlnAsp Lys Goes To Glu Ile Val Ile Gly Asp To Gly Arg Phe Arg Gln

485 490 495485 490 495

Ile Ile Thr Asn Leu vai Gly Asn Ser Val Lys Phe Thr Glu Arg Gly500 505 510Ile Ile Thr Asn Leu Goes Gly Asn Ser Val Lys Phe Thr Glu Arg Gly500 505 510

His Ile Phe Val Lys vai His Leu Ala Glu Asn ser Lys Val ser MetHis Ile Phe Val Lys Goes His Leu Wing Glu Asn Be Lys Val Be Met

515 520 525515 520 525

Asp Ser Glu Tyr Val Asn Gly Ile ser Asp Ser Gly Leu Phe vai Leu530 535 540Asp Be Glu Tyr Val Asn Gly Ile Be Asp Be Gly Leu Phe Goes Leu530 535 540

Asp Gly Arg Glu Phe Gln Thr Leu ser Gly Arg Glu Ala Ala Asp Asp545 550 555 560Asp Gly Arg Glu Phe Gln Thr Read to be Gly Arg Glu Wing Wing Asp Asp545 550 555 560

Gln Asn Ser Trp Asp Asn Phe Lys His Leu Ile Ala Asp Asp Asn Phe565 570 575Gln Asn Be Trp Asp Asn Phe Lys His Leu Ile Wing Asp Asp Asn Phe565 570 575

Gln Ser Asn Ala Ala Ser Asn Asn ser Ala Val Thr Asn Lys Gly Cys580 585 590Gln Be Asn Wing Ward Be Asn Asn Be Wing Val Thr Asn Lys Gly Cys580 585 590

Asp His vai Thr Leu Met vai Ser vai Glu Asp Thr Gly Ile Gly IleAsp His Goes Thr Leu Met Will Be Goes Glu Asp Thr Gly Ile Gly Ile

595 600 605595 600 605

Leu Leu His Ala Gln Asn Arg Val Phe Thr Pro Phe Met Gln Ala Asp610 615 620Leu Leu His Wing Gln Asn Arg Val Phe Thr Pro Phe Met Gln Wing Asp610 615 620

Ser Ser Thr Ser Arg Asn Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile625 630 635 640Be Ser Thr Be Arg Asn Tyr Gly Gly Thr Gly Ile Gly Read Ser Be Ile625 630 635 640

Ser Lys Cys Leu Val Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Ser645 650 655Ser Lys Cys Leu Val Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Ser645 650 655

Arg Pro Gln Ile Gly Ser Thr Phe Ser Phe Thr Ala Val Phe Gly LysArg Pro Gln Ile Gly Be Thr Phe Be Phe Thr Wing Val Phe Gly Lys

660 665 670660 665 670

Cys Lys Lys Asn Ser Met Asn Asp Met Lys Lys Pro Asn Ser Glu GluCys Lys Lys Asn Be Met Asn Asp Met Lys Lys Pro Asn Be Glu Glu

675 680 685675 680 685

Leu Pro Pro Ser Phe Arg Gly Met Lys Ala Ile Val vai Asp ser Lys690 695 700Leu Pro Pro Be Phe Arg Gly Met Lys Wing Ile Val Will Asp Be Lys690 695 700

His Val Arg Ala Ser Val Thr Arg Tyr His Leu Lys Arg Leu Gly Ile705 710 715 720His Val Arg Wing Be Val Thr Arg Tyr His Leu Lys Arg Leu Gly Ile705 710 715 720

Ile Val Glu vai Thr Asn ser Ile Asn Met Ala Ala Ser Leu Phe Arg725 730 735Ile Val Glu Will Thr Asn Be Ile Asn Met Wing Wing Be Leu Phe Arg725 730 735

Glu Asn Gly Ser Thr Leu Pro Arg Asn Thr Ile Leu Pro Asp Met Ile740 745 750Glu Asn Gly Ser Thr Thru Asu Arg Asn Thr Ile Leu Pro Asp Met Ile740 745 750

Leu vai Glu Lys Asp Ile Leu Asn Ser Asp Glu Glu Cys Gly Ile Ile755 760 765Leu Goes Glu Lys Asp Ile Leu Asn Be Asp Glu Glu Cys Gly Ile Ile755 760 765

His His Leu Asn Trp Lys Pro Asn Gly ser ser Val Lys Phe Pro LysHis His Leu Asn Trp Lys Pro Asn Gly Be Val Lys Phe Pro Lys

770 775 780770 775 780

Leu Ile Leu Leu Ala Thr Asn Ile Ala Thr Ala Glu Leu Asp Lys Ala785 790 795 800Leu Ile Leu Leu Wing Thr Thr Asn Ile Wing Thr Thr Glu Leu Asp Lys Ala785 790 795 800

Arg Ala Ala Gly Phe Ala Asp Thr vai Ile Met Lys Pro Leu Arg AlaArg Wing Gly Phe Wing Asp Thr Go Ile Met Lys Pro Read Arg Wing

805 810 815805 810 815

Thr Met Val Ala Ala Cys Leu Gln Gln Val Leu Gly Val Lys Asn Gln820 825 8B0Thr Met Val Wing Cys Wing Read Gln Gln Val Read Gly Val Lys Asn Gln820 825 8B0

Arg Arg Pro Asn Gly Ser Ala Phe Leu Gln Ser Leu Leu Cys Gly Lys835 840 845Arg Arg Pro Asn Gly Be Ala Phe Read Gln Be Read Leu Cys Gly Lys835 840 845

Arg Ile Leu Ile Val Asp Asp Asn Arg Val Asn Arg Arg vai Ala AlaArg Ile Leu Ile Val Asp Asp Asn Arg Val Asn Arg Arg goes Wing Wing

850 855 860850 855 860

Gly Ala Leu Lys Lys Phe Gly Ala Asp vai Glu Cys Ala Asp Ser Gly865 870 875 880Gly Wing Read Lys Lys Phe Gly Wing Asp Goes Glu Cys Wing Asp Ser Gly865 870 875 880

Lys Ser Ala Leu Lys Leu Leu Gln Leu Pro His Asn Phe Asp Ala CysLys Ser Ala Leu Lys Leu Leu Gln Leu Pro His Asn Phe Asp Cys Wing

885 890 895885 890 895

Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala Thr Arg900 905 910Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Wing Thr Arg900 905 910

Arg lie Arg Thr Met Glu Val Glu Ala Asn Lys Gly Gly Leu Ser Ala915 920 925Arg lie Arg Thr Met Glu Val Glu Wing Asn Lys Gly Gly Leu Ser Ala915 920 925

Thr Glu Gly Lys Arg Pro Ile Pro Ile Leu Ala Met Thr Ala Asp ValGlu Thr Gly Lys Arg Pro Ile Pro Ile Leu Wing Met Thr Wing Asp Val

930 935 940930 935 940

Ile His Ala Thr Tyr Glu Glu Cys Leu Lys Cys Gly Met Asn Gly Tyr945 950 955 960Ile His Wing Thr Tyr Glu Glu Cys Read Lys Cys Gly Met Asn Gly Tyr945 950 955 960

vai Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr Lys Glu Val Ala Argwill be Lys Pro Phe Glu Glu Glu Asn Leu Tyr Lys Glu Val Wing Arg

965 970 975965 970 975

Phe Phe Lys Lys Pro980Phe Phe Lys Lys Pro980

<210> 15<211> 993<212> PRT<213> Lotus corniculatus<220><210> 15 <211> 993 <212> PRT <213> Lotus corniculatus <220>

<221> Mutation<222> (266)..(266)<223> X: is Xaa (any anrino acid other than Leu)<220><221> Mutation <222> (266) .. (266) <223> X: is Xaa (any anrino acid other than Leu) <220>

<221> misc_feature<222> (266)..(266)<221> misc_feature <222> (266) .. (266)

<223> Xaa can be any natural!y occurring amino acid<400> 15<223> Xaa can be any natural! And occurring amino acid <400> 15

Met Gly Leu Gly Phe Lys Met Gln Gln Ser His His Pro Val Ala Leu1 5 10 15Met Gly Leu Gly Phe Lys Met Gln Gln Be His His Pro Val Wing Leu1 5 10 15

Lys Leu His Glu Gln Ala Gly Ser Gln Arg Lys Phe Thr Phe Ile Gln20 25 30Lys Read His Glu Gln Wing Gly Be Gln Arg Lys Phe Thr Phe Ile Gln20 25 30

15 Asn Phe Arg Asn Trp Phe Leu Pro Leu Leu Phe vai Trp Phe Ile Val35 40 4515 Asn Phe Arg Asn Trp Phe Leu Pro Leu Leu Phe Goes Trp Phe Ile Val35 40 45

Met Ala Ala Phe Gly Ala Cys Ile Tyr His Lys Met Asp Ala Glu ThrMet Asp Wing Phe Gly Cys Wing Ile Tyr His Lys Met Asp Wing Glu Thr

50 55 6050 55 60

Lys vai Arg Arg Lys Glu vai Leu Gly Ser Leu Cys Asp Gln Arg Ala20 65 70 75 80Lys Goes Arg Arg Lys Glu Goes Read Gly Be Read Cys Asp Gln Arg Wing20 65 70 75 80

Arg Met Leu Gln Asp Gln Phe Ser vai Ser Val Asn His Val His AlaArg Met Read Gln Asp Gln Phe Ser Will Be Val Asn His Val His Wing

85 90 9585 90 95

Leu Ala Ile Leu vai Ser Thr Phe His Tyr Tyr Arg Asn Thr Ser Ala100 105 110Leu Ala Ile Leu Will Be Thr Phe His Tyr Tyr Arg Asn Thr Be Ala100 105 110

25 Ile Asp Gln Glu Thr Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe Glu115 120 12525 Ile Asp Gln Glu Thr Phe Wing Glu Tyr Thr Wing Arg Thr Phe Wing Glu115 120 125

Arg Pro Leu Met ser Gly Val Ala Tyr Ala Gln Arg Val vai His serArg Pro Read Met Be Gly Val Wing Tyr Wing Gln Arg Val Goes His Be

130 135 140130 135 140

Glu Arg Glu Arg Phe Glu Lys Gln His Gly Trp vai Ile Lys Thr Met30 145 150 155 160Glu Arg Glu Arg Phe Glu Lys Gln His Gly Trp Goes Ile Lys Thr Met30 145 150 155 160

Glu Arg Val Pro Ser Gly Val Arg Asp Glu Tyr Ala Ala vai Ile Phe165 170 175Ala Gln Glu Thr vai Ser Tyr Leu Glu Ser Ile Asp Met Met ser GlyGlu Arg Val Pro To Be Gly Val Arg Asp Glu Tyr Wing Ward Go Ile Phe165 170 175Ala Gln Glu Thr Go To Be Tyr Leu Glu Be Ile Asp Met Met Be Gly

180 185 190180 185 190

Glu Glu Asp Arg Glu Asn Ile Leu Arg Ala Arg Ala Thr Gly Lys Ala195 200 205Glu Glu Asp Arg Glu Asn Ile Leu Arg Wing Arg Wing Arg Wing Thr Gly Lys Wing195 200 205

Val Leu Thr Ser Pro Phe Arg Leu Leu Asp Ser His His Leu Gly Val210 215 220Val Leu Thr Be Pro Phe Arg Leu Read Asp Be His His Leu Gly Val210 215 220

Val Leu Thr Phe Pro Val Tyr Lys Ser Lys Leu Pro Pro Glu Pro Thr225 230 235 240Val Leu Thr Phe Pro Val Tyr Lys Ser Lys Leu Pro Pro Glu Pro Thr225 230 235 240

Thr Glu Glu Val Ile Lys Ala Ile Ala Gly Tyr Ile Gly Gly Ser Phe 245 250 255Thr Glu Glu Val Ile Lys Wing Ile Wing Gly Tyr Ile Gly Gly Ser Phe 245 250 255

Asp Val Glu Ser Leu Val Glu Asn Leu Xaa Gly Gln Leu Ala Gly AsnAsp Val Glu Ser Leu Val Glu Asn Leu Xaa Gly Gln Leu Wing Gly Asn

260 265 270260 265 270

Gln Ala Ile Leu vai Lys Val Tyr Asp Ile Thr Asn ser ser Asp Pro275 280 285Gln Wing Ile Leu Will Lys Val Tyr Asp Ile Thr Asn Be Being Asp Pro275 280 285

Leu Ile Met Tyr Gly Ser Gln Tyr Glu Glu Gly Asp Met Ser Leu Val290 295 300Leu Ile Met Tyr Gly Ser Gln Tyr Glu Glu Gly Asp Met Ser Leu Val290 295 300

His Glu Ser Lys Leu Asp Phe Gly Asp Pro Tyr Arg Lys His His Met305 310 315 320His Glu Ser Lys Read Asp Phe Gly Asp Pro Tyr Arg Lys His His Met305 310 315 320

Ile Cys Arg Tyr His Gln Gln Ala Pro Thr Asn Trp Ile Ala Tyr Thr 325 330 335Ile Cys Arg Tyr His Gln Gln Wing Pro Thr Asn Trp Ile Cys Wing Tyr Thr 325 330 335

Thr Ala Phe Leu Phe Phe Val Ile Leu Cys Leu vai Gly Tyr Ile LeuThr Ala Phe Leu Phe Phe Val Ile Leu Cys Leu goes Gly Tyr Ile Leu

340 345 350340 345 350

Tyr Ala Ala Gly Thr His Ile Val Lys vai Glu Asp Asp Tyr Asn Ala355 360 365Tyr Ala Wing Gly Thr His Ile Val Lys Goes Glu Asp Asp Tyr Asn Ala355 360 365

Met Gln Asp Leu Lys vai Lys Ala Glu Ala Ala Asp Ile Ala Lys Ser370 375 380Met Gln Asp Read Lys Go Lys Wing Glu Wing Wing Asp Ile Wing Lys Ser370 375 380

Gln Phe Leu Ala Thr Val Ser His Glu Ile Arg Thr Pro Met Asn Gly385 390 395 400Gln Phe Read Wing Thr Val Be His Glu Ile Arg Thr Pro Met Asn Gly385 390 395 400

Ile Leu Gly Met Leu Gly Leu Leu Leu Arg Thr Glu Leu Ser Ser Thr 405 410 415Ile Leu Gly Met Leu Gly Leu Leu Read Leu Arg Thr Glu Leu Be Ser Thr 405 410 415

Gln Arg Asp Tyr Ala Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu Ile420 425 430Ala Leu Ile Asn Glu vai Leu Asp Arg Ala Lys Ile Glu Ala Gly LysGln Arg Asp Tyr Wing Gln Thr Wing Gln Cys Wing Gly Lys Wing Leu Ile420 425 430Ala Leu Ile Asn Glu Goes To Read Asp Arg Wing Lys Ile Glu Wing Gly Lys

435 440 445435 440 445

Leu Glu Leu Glu Ala vai Pro Phe Asp Leu Arg Ser Ile Leu Asp Asp450 455 460Leu Glu Leu Glu Wing goes to Phe Asp Leu Arg Ser Ile Leu Asp Asp450 455 460

vai Leu Ser Leu Phe Ser Glu Lys Ser Arg His Lys Gly Leu Glu Leu465 470 475 480will Read Be Read Read Phe Be Glu Lys Be Arg His Lys Gly Leu Glu Leu465 470 475 480

Ala Val Phe Val Ser Asp Lys vai Pro Asp Ile vai Met Gly Asp ProVal Phe Val Wing Asp Lys Go Pro Asp Ile Go Met Gly Asp Pro

485 490 495485 490 495

Gly Arg Phe Arg Gln Ile Val Thr Asn Leu Val Gly Asn Ser Val Lys 500 505 510Gly Arg Phe Arg Gln Ile Val Thr Asn Read Val Gly Asn Ser Val Lys 500 505 510

Phe Thr Glu Arg Gly His Ile Phe vai Lys vai His Leu Ala Glu LysPhe Thr Glu Arg Gly His Ile Phe Goes Lys Goes His Leu Wing Glu Lys

515 520 525515 520 525

Arg Gln Cys Thr Met Asn Gly Lys Cys Glu Thr Phe Leu Asn Gly Gly530 535 540Arg Gln Cys Met Thr Asn Gly Lys Cys Glu Thr Phe Read Asn Gly Gly530 535 540

Cys Asp Asp Val Leu His Val Ser Gly Ser Tyr Asn Leu Lys Thr Leu545 550 555 560Cys Asp Asp Val Leu His Val Ser Gly Ser Tyr Asn Leu Lys Thr Leu545 550 555 560

Ser Gly Tyr Glu Ala Ala Asp Glu Arg Asn Ser Trp Asp Asn Phe LysBe Gly Tyr Glu Wing Wing Asp Glu Arg Asn Be Trp Asp Asn Phe Lys

565 570 575565 570 575

His His Ile Ala Asp Glu Glu Phe Phe Phe Asp Ala Ser Val Lys Lys 580 585 590His His Ile Wing Asp Glu Glu Phe Phe Phe Asp Wing Be Val Lys Lys 580 585 590

Leu Ala Ser Ser Glu Ser Tyr Glu Gln vai Thr Leu Met Val Ser ValLeu Wing Be Ser Glu Ser Tyr Glu Gln Go Thr Leu Met Val Ser Val

595 600 605595 600 605

Glu Asp Thr Gly Ile Gly Ile Ser Phe Ser Ala Gln Asp Ser Ile Phe610 615 620Glu Asp Thr Gly Ile Gly Ile Be Phe Be Wing Gln Asp Be Ile Phe610 615 620

Met Pro Phe Val Gln Ala Asp Ser Ser Thr Ser Arg Asn Tyr Gly Gly625 630 635 640Met Pro Phe Val Gln Wing Asp Be Be Thr Be Arg Asn Tyr Gly Gly625 630 635 640

Thr Gly Ile Gly Leu Ser Ile Ser Lys Cys Leu Val Glu Leu Met GlyThr Gly Ile Gly Leu Be Ile Be Lys Cys Leu Val Glu Leu Met Gly

645 650 655645 650 655

Gly Gln Ile Asn Phe Ile Ser Arg Pro Gln Val Gly Ser Thr Phe Ser 660 665 670Gly Gln Ile Asn Phe Ile Be Arg Pro Gln Val Gly Be Thr Phe Ser 660 665 670

Phe Thr Ala Asp Phe Gly Thr Phe Lys Lys Asn ser Thr Thr Asp Met675 680 685Lys Lys Leu Asn Phe Glu Asp Leu Pro Ser Ser Phe Arg Gly Leu LysPhe Thr Wing Asp Phe Gly Thr Phe Lys Lys Asn Be Thr Thr Asp Met675 680 685Lys Lys Leu Asn Phe Glu Asp Leu Pro To Be Phe Arg Gly Leu Lys

690 695 700690 695 700

Ala Ile vai vai Asp Gly Lys Pro Val Arg Ala Ala vai Thr Arg Tyr705 710 715 720Wing Ile Goes Go Asp Gly Lys Pro Val Arg Wing Wing Goes Thr Arg Tyr705 710 715 720

His Leu Lys Arg Leu Gly Ile Gln Ala Lys vai Ala Ile ser Ile AsnHis Leu Lys Arg Leu Gly Ile Gln Wing Lys Go Wing Ile Be Ile Asn

725 730 735725 730 735

Lys Ala vai Ser Leu Cys Gly Lys Asn Gly Ser Leu Thr Ser Ala LeuLys Ala Will Be Leu Cys Gly Lys Asn Gly Be Leu Thr Be Ala Leu

740 745 750740 745 750

Phe Gln Pro Asp Ile Ile Phe Val Glu Lys Asp Ser Trp Val Ser Gly755 760 765Phe Gln Pro Asp Ile Ile Phe Val Glu Lys Asp Ser Trp Val Ser Gly755 760 765

Glu Asp Gly Gly Ile Phe Asn Ala Phe Lys Met Pro Gln Met Ile LeuGlu Asp Gly Gly Ile Phe Asn Ala Phe Lys Met Pro Gln Met Ile Leu

770 775 780770 775 780

Leu Ala Thr Asn Ile Cys Asn Ala Glu Phe Asp Lys Ala Lys Ala Ala785 790 795 800Leu Wing Thr Asn Ile Cys Asn Wing Glu Phe Asp Lys Wing Lys Wing Wing785 790 795 800

Gly Phe Ser Asp Thr Val Ile Met Lys Pro Leu Arg Ala Ser Met LeuGly Phe Ser Asp Thr Val Ile Met Lys Pro Leu Arg Wing Be Met Leu

805 810 815805 810 815

Ala Ala Cys Leu Gln Gln vai Phe Gly Thr Gly Lys Thr Arg Gln PheWing Cys Wing Read Gln Gln Go Phe Gly Thr Gly Lys Thr Arg Gln Phe

820 825 830820 825 830

Gly Lys Asp Met Ser Asn Gly Ser Ser vai Arg Ser Leu Leu Cys Gly835 840 845Gly Lys Asp Met Be Asn Gly Be Be Will Arg Be Read Leu Cys Gly835 840 845

Lys Lys Ile Leu Val Val Asp Asp Asn Leu vai Asn Arg Arg Val AlaLys Lys Ile Leu Val Val Asp Asp Asn Leu Goes Asn Arg Arg Val Wing

850 855 860850 855 860

Ala Gly Ala Leu Lys Asn Phe Gly Ala Asp vai Lys Cys Ala Ala Ser865 870 875 880Wing Gly Wing Read Lys Asn Phe Gly Wing Asp Goes Lys Cys Wing Ser865 870 875 880

Gly Lys Ala Ala Leu Glu Met Leu Gln Tyr Pro His Asp Phe Asp AlaGly Lys Wing Wing Read Glu Met Read Gln Tyr Pro His Asp Phe Asp Wing

885 890 895885 890 895

Cys Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala ThrCys Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala Thr

900 905 910900 905 910

Arg Arg Ile Arg Met Met Glu Arg Glu Ala Ser Glu Gln Leu Lys ser915 920 925Arg Arg Ile Arg Met Met Glu Arg Glu Wing Be Glu Gln Leu Lys ser915 920 925

Glu Ser Gly Glu Glu Asn Gly Lys Lys Ser Glu Phe His Met Pro Ile930 935 940Leu Ala Met Thr Ala Asp vai Ile His Ala Thr Tyr Asp Lys Cys Leu945 950 955 960Glu Be Gly Glu Glu Asn Gly Lys Lys Be Glu Phe His Met Pro Ile930 935 940Leu Wing Met Thr Wing Asp Go Ile His Wing Thr Tyr Asp Lys Cys Leu945 950 955 960

Asn Cys Gly Met Asp Gly Tyr Val Ser Lys Pro Phe Glu Glu Glu Asn965 970 975Asn Cys Gly Met Asp Gly Tyr Val Ser Lys Pro Phe Glu Glu Glu Asn965 970 975

Leu Tyr Gln Ala Val Ala Lys Phe Phe Lys Ser Lys Pro Ala Ser Asp980 985 990Read Tyr Gln Wing Val Wing Lys Phe Phe Lys Ser Lys Pro Wing Ser Asp980 985 990

SerTo be

<210> 16<211> 1003<212> PRT<210> 16 <211> 1003 <212> PRT

<213> Medicago truncatula<220><213> Medicago truncatula <220>

<221> Mutation<222> (267)..(267)<221> Mutation <222> (267) .. (267)

<223> X = Xaa (any amino aicd other than Leu)<220><223> X = Xaa (any amino aicd other than Leu) <220>

<221> misc_feature<222> (267)..(267)<223> Xaa can be any naturally occurring amino acid<400> 16<221> misc_feature <222> (267) .. (267) <223> Xaa can be any naturally occurring amino acid <400> 16

Met Gly Leu Leu Leu Lys Met Lys Met Gln Asn Gln His His Pro Leu1 5 10 15Met Gly Leu Leu Leu Lys Met Lys Met Gln Asn Gln His His Pro Leu1 5 10 15

Ala Ser Lys Leu Gln Glu Gln Thr Gly Asn Lys Arg Tyr Thr Phe Ile20 25 30Ala Ser Lys Leu Gln Glu Gln Thr Gly Asn Lys Arg Tyr Thr Phe Ile20 25 30

Gln Ala His Arg Ala Trp Leu Leu Lys Leu Met Phe Leu Trp Ile LeuGln Wing His Arg Wing Trp Leu Leu Lys Leu Met Phe Leu Trp Ile Leu

35 40 4535 40 45

Leu Met Ala Leu Ile Ser Arg Ile Ile Tyr Ser Lys Met Asp Val Gly50 55 60Leu Met Ala Leu Ile Be Arg Ile Ile Tyr Be Lys Met Asp Val Gly50 55 60

Thr Lys vai Arg Arg Lys Glu vai Leu Gly Ser Leu Cys Asp Gln Arg65 70 75 80Thr Lys Will Arg Arg Lys Glu Will Read Gly Be Read Cys Asp Gln Arg65 70 75 80

Ala Arg Met Leu Gln Asp Gln Phe Ser Val Ser Val Asn His Val His85 90 95Wing Arg Met Read Gln Asp Gln Phe Ser Val Ser Val Asn His Val His85 90 95

Ala Leu Ala Ile Leu Val Ser Thr Phe His Tyr Tyr Arg Asn Pro SerAla Leu Ala Ile Leu Val Be Thr Phe His Tyr Tyr Arg Asn Pro Ser

100 105 110100 105 110

Ala Ile Asp Gln Glu Thr Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe5 115 120 125Wing Ile Asp Gln Glu Thr Phe Wing Glu Tyr Thr Wing Arg Thr Wing Phe5 115 120 125

Glu Arg Pro Leu Leu Ser Gly Val Ala Tyr Ala Gln Arg Val Val AsnGlu Arg Pro Leu Read Ser Gly Val Wing Tyr Wing Gln Arg Val Val Asn

130 135 140130 135 140

Ser Glu Arg Glu Gln Phe Glu Lys Gln His Gly vai vai Ile Lys Thr145 150 155 160Be Glu Arg Glu Gln Phe Glu Lys Gln His Gly Goes Ile Lys Thr145 150 155 160

10 Met Glu Arg Glu Ala Ser Pro Val Arg Asp Glu Tyr Ala Pro Val Ile10 Met Glu Arg Glu Wing Be Pro Val Arg Asp Glu Tyr Wing Pro Val Ile

165 170 175165 170 175

Phe Ala Gln Glu Thr Val Ser Tyr Leu Glu Ser Ile Asp Met Met SerPhe Ala Gln Glu Thr Val Ser Tyr Leu Glu Ser Ile Asp Met Met Ser

180 185 190180 185 190

Gly Glu Glu Asp Arg Glu Asn Ile Met Arg Ala Arg Ala Thr Gly Lys15 195 200 205Gly Glu Glu Asp Arg Glu Asn Ile Met Arg Wing Arg Wing Wing Thr Gly Lys15 195 200 205

Ala Val Leu Thr Ser Pro Phe Arg Leu Leu Gly Ser His His Leu GlyWing Val Leu Thr Be Pro Phe Arg Leu Read Gly Be His His Leu Gly

210 215 220210 215 220

vai vai Leu Thr Phe Pro Val Tyr Lys Ser Lys Leu Pro Pro Asn Pro225 230 235 240go go Read Thr Phe Pro Val Tyr Lys Ser Lys Leu Pro Pro Asn Pro225 230 235 240

20 Thr Thr Glu Glu Leu Ile Lys Ala Thr Ala Gly Tyr vai Gly Gly Ser20 Thr Thr Glu Glu Leu Ile Lys Wing Thr Wing Gly Tyr will Gly Gly Ser

245 250 255245 250 255

Phe Asp Val Glu Ser Leu vai Glu Asn Leu Xaa Gly Gln Leu Ala GlyPhe Asp Val Glu Being Leu Goes Glu Asn Leu Xaa Gly Gln Leu Wing Gly

260 265 270260 265 270

His Gln Ala Ile Leu vai Asn vai Tyr Asp Val Thr Asn Ser Ser Asp25 275 280 285His Gln Wing Ile Leu Goes Asn Goes Tyr Asp Val Thr Asn Being Ser Asp25 275 280 285

Pro Leu Ile Met Tyr Gly Asn Gln Tyr Glu Glu Gly Asp vai Ser LeuPro Leu Ile Met Tyr Gly Asn Gln Tyr Glu Glu Gly Asp Will Be Leu

290 295 300290 295 300

Val His Glu Ser Lys Leu Asp Phe Gly Asp Pro Tyr Arg Lys His Gln305 310 315 320Val His Glu Ser Lys Read Asp Phe Gly Asp Pro Tyr Arg Lys His Gln305 310 315 320

30 Met Ile Cys Arg Tyr His Gln Lys Ala Pro Pro Asn Trp Thr Ala Leu30 Met Ile Cys Arg Tyr His Gln Lys Pro Pro Asn Trp Thr Wing Leu

325 330 335325 330 335

Ser Thr Ala lie Leu Phe Phe Val Ile Leu Leu Leu Ile Gly Tyr Ile340 345 350Ser Thr Ala lie Leu Phe Phe Val Ile Leu Leu Leu Ile Gly Tyr Ile340 345 350

Leu Tyr Gly Ala Gly Asn His Ile Val Lys vai Glu Asp Asp Phe His355 360 365Read Tyr Gly Wing Gly Asn His Ile Val Lys Goes Glu Asp Asp Phe His355 360 365

Glu Met Gln Glu Leu Lys Val Arg Ala Glu Ala Ala Asp vai Ala Lys370 375 380Glu Met Gln Glu Leu Lys Val Arg Wing Glu Wing Wing Asp goes Wing Lys370 375 380

Ser Gln Phe Leu Ala Thr Val Ser His Glu Ile Arg Thr Pro Met Asn385 390 395 400Be Gln Phe Leu Thr Wing Val Be His Glu Ile Arg Thr Pro Met Asn385 390 395 400

Gly Ile Leu Gly Met Leu Gly Leu Leu Leu Arg Thr Glu Leu Asn Ser405 410 415Gly Ile Leu Gly Met Leu Gly Leu Leu Leu Leu Arg Thr Glu Leu Asn Ser405 410 415

Thr Gln Arg Asp Tyr Ala Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu420 425 430Thr Gln Arg Asp Tyr Gln Wing Thr Gln Wing Cys Wing Gly Lys Wing Leu420 425 430

Ile Ala Leu Ile Asn Glu Val Leu Asp Arg Ala Lys Ile Glu Ala Gly435 440 445Ile Wing Leu Ile Asn Glu Val Leu Asp Arg Wing Lys Ile Glu Wing Gly435 440 445

Lys Leu Glu Leu Glu Ala vai Pro Phe Asp Leu Arg Ser Ile Leu Asp450 455 460Lys Leu Glu Leu Glu Wing Goes To Phe Asp Leu Arg Be Ile Leu Asp450 455 460

Asp Val Leu ser Leu Phe Ser Glu Lys ser Arg His Lys Gly Leu Glu465 470 475 480Asp Val Leu Be Leu Phe Be Glu Lys Be Arg His Lys Gly Leu Glu465 470 475 480

Leu Ala Val Phe vai ser Asp Lys Val Pro Asp Ile vai Met Gly Asp485 490 495Leu Wing Val Phe Will Be Asp Lys Val Pro Asp Ile Will Met Gly Asp485 490 495

Pro Gly Arg Phe Arg Gln Ile Val Thr Asn Leu Val Gly Asn Ser Val500 505 510Pro Gly Arg Phe Arg Gln Ile Val Thr Asn Read Val Gly Asn Ser Val500 505 510

Lys Phe Thr Glu Arg Gly His Ile Phe Val Lys vai His Leu Ser Glu515 520 525Lys Phe Thr Glu Arg Gly His Ile Phe Val Lys Goes His Leu Being Glu515 520 525

Asn Arg Lys Pro Val Thr Asn Gly Lys His Glu Thr Tyr Arg Asn Gly530 535 540Asn Arg Lys Pro Val Asn Gly Lys His Glu Thr Tyr Arg Asn Gly530 535 540

Gly Ser Glu Glu vai Val His Ala Ser Gly Gly Tyr Asn Leu Lys Thr545 550 555 560Gly Ser Glu Glu Goes Val His Wing Ser Gly Gly Tyr Asn Leu Lys Thr545 550 555 560

Leu Ser Gly Tyr Glu Ala Ala Asp Glu Arg Asn Asn Trp Asp Asn Phe565 570 575Leu Ser Gly Tyr Glu Wing Wing Asp Glu Arg Asn Asn Asp Trp Asp Ashe Phe565 570 575

Asn His Leu Ile Ala Asp Glu Glu Phe Phe Cys Asp Ala ser Thr Lys580 585 590Asn His Leu Ile Wing Asp Glu Glu Phe Phe Cys Asp Wing Be Thr Lys580 585 590

Lys vai Ala ser Asn Glu Phe Tyr Glu Gln Val Thr Leu Met Val Cys595 600 605Lys will Ala be Asn Glu Phe Tyr Glu Gln Val Thr Leu Met Val Cys595 600 605

Val Glu Asp Thr Gly Ile Gly Ile Pro Phe Ser Ala Gln Asp Arg IleVal Glu Asp Thr Gly Ile Gly Ile Pro Phe Ser Wing Gln Asp Arg Ile

610 615 620610 615 620

Phe Met Pro Phe Val Gln Ala Asp Ser Ser Thr Ser Arg Asn Tyr Gly625 630 635 640Phe Met Pro Phe Val Gln Wing Asp Be Be Thr Be Arg Asn Tyr Gly625 630 635 640

Gly Thr Gly Ile Gly Leu Ser Ile Ser Lys Cys Leu Val Glu Leu MetGly Thr Gly Ile Gly Leu Be Ile Be Lys Cys Leu Val Glu Leu Met

645 650 655645 650 655

Gly Gly Gln Ile Asn Phe lie Ser Arg Pro Gln Val Gly Ser Thr Phe660 665 670Gly Gly Gln Ile Asn Phe lie Ser Arg Pro Gln Val Gly Ser Thr Phe660 665 670

Ser Phe Thr Ala Asp Phe Gly Ile Phe Lys Lys Asn Pro Ile Thr Glu675 680 685Ser Phe Thr Wing Asp Phe Gly Ile Phe Lys Lys Asn Pro Ile Thr Glu675 680 685

vai Lys Lys vai Asn Tyr Glu Asp Leu Pro Ser Ser Phe Arg Gly Leugo Lys Lys go Asn Tyr Glu Asp Leu Pro Be Ser Phe Arg Gly Leu

690 695 700690 695 700

Lys Ala Val Val vai Asp Gly Lys Pro Val Arg Ala Ala Val Thr Arg705 710 715 720Lys Wing Val Val Go Asp Gly Lys Pro Val Arg Wing Wing Val Thr Arg705 710 715 720

Tyr His Leu Lys Arg Leu Gly Ile Gln vai Lys Val Ala Asn Ala IleTyr His Leu Lys Arg Leu Gly Ile Gln Goes Lys Val Wing Asn Wing Ile

725 730 735725 730 735

Asn Lys Ala Val Ser Leu Cys Gly Lys Asn Gly Ala Ser Ser Thr Gly740 745 750Asn Lys Wing Val Be Read Cys Gly Lys Asn Lys Wing Be Ser Thr Thr Gly740 745 750

Leu Phe Gln Pro Asp Ile Ile Phe Val Glu Lys Asp Ser Trp Val Cys755 760 765Leu Phe Gln Pro Asp Ile Ile Phe Val Glu Lys Asp Ser Trp Val Cys755 760 765

Gly Glu Asp Gly Ile Phe Ser Val Arg Gln Leu Asp Trp Lys Gln AsnGly Glu Asp Gly Ile Phe Ser Val Arg Gln Read Asp Trp Lys Gln Asn

770 775 780770 775 780

Gly His Ile Phe Lys Met Pro Gln Met Ile Leu Leu Ala Thr Asn Ile785 790 795 800Gly His Ile Phe Met Lys Pro Gln Met Ile Read Leu Wing Thr Asn Ile785 790 795 800

Ser Asn Asp Glu Phe Asp Lys Ala Lys ser Ala Gly Phe ser Asp ThrBe Asn Asp Glu Phe Asp Lys Wing Lys Be Wing Gly Phe Be Asp Thr

805 810 815805 810 815

vai Ile Met Lys Pro Leu Arg Ala Ser Met Val Gly Ala Cys Leu Gln820 825 830Go Ile Met Lys Pro Read Arg Wing Be Met Val Gly Cys Wing Read Gln820 825 830

Gln vai Leu Gly Thr Gly Lys Lys Arg Gln Leu Gly Lys Glu Met Pro835 840 845Gln Will Read Gly Thr Gly Lys Lys Arg Gln Read Gly Lys Glu Met Pro835 840 845

Asn Gly Ser Thr Ser Val Arg Ser Leu Leu Phe Gly Lys Lys Ile Leu850 855 860Asn Gly Be Thr Be Val Arg Be Read Leu Phe Gly Lys Lys Ile Leu850 855 860

val Val Asp Asp Asn Val Val Asn Arg Arg Val Ala Ala Gly Ala Leu865 870 875 880val Val Asp Asp Asn Val Val Asn Arg Arg Val Wing Gly Wing Leu865 870 875 880

Lys Asn Phe Gly Ala Asp Val Lys Cys Ala Asp Ser Gly Lys Ala Ala885 890 895Lys Asn Phe Gly Wing Asp Val Lys Cys Wing Asp Ser Gly Lys Wing Ala885 890 895

Leu Glu Met Leu Gln Phe Pro His Lys Phe Asp Ala Cys Phe Met Asp900 905 910Read Glu Met Read Le Gln Phe Pro His Lys Phe Asp Cys Wing Phe Met Asp900 905 910

Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala Thr Arg Arg Ile Arg915 920 925Ile Gln Met Pro Glu Met Asp Gly Phe Glu Wing Thr Arg Arg Ile Arg915 920 925

Glu Met Glu Arg Thr Ala Asn Glu Glu Thr Asn Ser Glu Cys Gly Glu930 935 940Glu Met Glu Arg Thr Asn Wing Glu Glu Thr Asn Be Glu Cys Gly Glu930 935 940

Arg Lys Ser Glu Phe His Leu Pro Ile Leu Ala Met Thr Ala Asp Val945 950 955 960Arg Lys Ser Glu Phe His Leu Pro Ile Leu Wing Met Thr Wing Asp Val945 950 955 960

Ile His Ala Thr Tyr Glu Glu Cys Leu Lys Cys Gly Met Asp Gly Tyr965 970 975Ile His Wing Thr Tyr Glu Glu Cys Read Lys Cys Gly Met Asp Gly Tyr965 970 975

Val Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr Gln Ala vai Ala Lys980 985 990Val Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr Gln Wing Go Wing Lys980 985 990

Phe Phe Gln Thr Lys Pro Thr Ser Val Asp Ser995 1000Phe Phe Gln Thr Lys Pro Thr Ser Val Asp Ser995 1000

<210> 17<211> 1080<212> PRT<213> Arabidopsis thaliana<220><221> Mutation<222> (356)..(356)<223> X= xaa (any amino acid other than Leucine)<220><221> misc_feature<222> (356)..(356)<223> Xaa can be any naturalIy occurring amino acid<400> 17Met Arg Arg Asp Phe Val Tyr Asn Asn Asn Ala Met Phe Asn Pro Leu1 5 10 15<210> 17 <211> 1080 <212> PRT <213> Arabidopsis thaliana <220> <221> Mutation <222> (356) .. (356) <223> X = xaa (any amino acid other than Leucine) < 220> <221> misc_feature <222> (356) .. (356) <223> Xaa can be any naturalIy occurring amino acid <400> 17Met Arg Arg Asp Phe Val Tyr Asn Asn Ala Met Phe Asn Pro Leu1 5 10 15

Thr Thr His Tyr Ser Ser Asp Met Asn Trp Ala Leu Asn Asn His Gln20 25 30Thr Thr His Tyr Being Ser Asp Met Asn Trp Wing Read Asn Asn His Gln20 25 30

Glu Glu Glu Glu Glu Pro Arg Arg Ile Glu Ile Ser Asp Ser Glu Ser35 40 45Glu Glu Glu Glu Glu Pro Arg Arg Ile Glu Ile Ser Asp Ser Glu Ser35 40 45

Leu Glu Asn Leu Lys Ser Ser Asp Phe Tyr Gln Leu Gly Gly Gly GlyLeu Glu Asn Leu Lys Being Ser Asp Phe Tyr Gln Leu Gly Gly Gly Gly

50 55 6050 55 60

Ala Leu Asn Ser Ser Glu Lys Pro Arg Lys Ile Asp Phe Trp Arg Ser65 70 75 80Wing Read Asn Be Glu Lys Pro Arg Lys Ile Asp Phe Trp Arg Ser65 70 75 80

Gly Leu Met Gly Phe Ala Lys Met Gln Gln Gln Gln Gln Leu Gln HisGly Leu Met Gly Phe Ala Lys Met Gln Gln Gln Gln Gln Glu Leu Gln His

85 90 9585 90 95

Ser Val Ala vai Lys Met Asn Asn Asn Asn Asn Asn Asp Leu Met Gly100 105 110Ser Val Wing goes Lys Met Asn Asn Asn Asn Asn Asn Asn Asp Leu Met Gly100 105 110

Asn Lys Lys Gly Ser Thr Phe Ile Gln Glu His Arg Ala Leu Leu Pro115 120 125Asn Lys Lys Gly Be Thr Phe Ile Gln Glu His Arg Wing Read Leu Pro115 120 125

Lys Ala Leu Ile Leu Trp Ile Ile Ile Val Gly Phe Ile Ser Ser GlyLys Wing Leu Ile Leu Trp Ile Ile Ile Val Gly Phe Ile Ser Ser Gly

130 135 140130 135 140

Ile Tyr Gln Trp Met Asp Asp Ala Asn Lys Ile Arg Arg Glu Glu Val145 150 155 160Ile Tyr Gln Trp Met Asp Asp Wing Asn Lys Ile Arg Arg Glu Glu Val145 150 155 160

Leu Val Ser Met Cys Asp Gln Arg Ala Arg Met Leu Gln Asp Gln PheLeu Val Ser Met Cys Asp Gln Arg Wing Arg Met Leu Gln Asp Gln Phe

165 170 175165 170 175

Ser Val ser Val Asn His vai His Ala Leu Ala Ile Leu vai Ser Thr180 185 190Ser Val be Val Asn His will His Wing Leu Wing Ile Leu will Be Thr180 185 190

Phe His Tyr His Lys Asn Pro Ser Ala Ile Asp Gln Glu Thr Phe Ala195 200 205Phe His Tyr His Lys Asn Pro Be Ala Ile Asp Gln Glu Thr Phe Ala195 200 205

Glu Tyr Thr Ala Arg Thr Ala Phe Glu Arg Pro Leu Leu Ser Gly ValGlu Tyr Thr Wing Arg Thr Wing Phe Glu Arg Pro Read Leu Ser Gly Val

210 215 220210 215 220

Ala Tyr Ala Glu Lys Val Val Asn Phe Glu Arg Glu Met Phe Glu Arg225 230 235 240Tyr Wing Glu Wing Lys Val Val Asn Phe Glu Arg Glu Met Phe Glu Arg225 230 235 240

Gln His Asn Trp Val Ile Lys Thr Met Asp Arg Gly Glu Pro Ser Pro245 250 255Val Arg Asp Glu Tyr Ala Pro Val Ile Phe Ser Gln Asp Ser Val SerGln His Asn Trp Val Ile Lys Thr Met Asp Arg Gly Glu Pro Ser Pro245 250 255Val Arg Asp Glu Tyr Wing Pro Val Ile Phe Ser Gln Asp Ser Val Ser

260 265 270260 265 270

Tyr Leu Glu Ser Leu Asp Met Met Ser Gly Glu Glu Asp Arg Glu Asn275 280 285Tyr Read Glu Be Read Asp Met Met Be Gly Glu Glu Asp Arg Glu Asn275 280 285

Ile Leu Arg Ala Arg Glu Thr Gly Lys Ala vai Leu Thr ser Pro Phe290 295 300Ile Leu Arg Wing Arg Glu Thr Gly Lys Wing Go Leu Thr be Pro Phe290 295 300

Arg Leu Leu Glu Thr His His Leu Gly Val Val Leu Thr Phe Pro Val305 310 315 320Arg Leu Leu Glu Thr His His Leu Gly Val Val Leu Thr Phe Pro Val305 310 315 320

Tyr Lys Ser Ser Leu Pro Glu Asn Pro Thr vai Glu Glu Arg Ile Ala 325 330 335Tyr Lys Being Serve Leu Pro Glu Asn Pro Thr Goes Glu Glu Arg Ile Wing 325 330 335

Ala Thr Ala Gly Tyr Leu Gly Gly Ala Phe Asp Val Glu Ser Leu ValWing Thr Wing Wing Gly Tyr Leu Gly Gly Wing Phe Asp Val Glu Ser Leu Val

340 345 350340 345 350

Glu Asn Leu Xaa Gly Gln Leu Ala Gly Asn Gln Ala Ile Val Val His355 360 365Glu Asn Leu Xaa Gly Gln Leu Wing Gly Asn Gln Wing Ile Val Val His355 360 365

Val Tyr Asp Ile Thr Asn Ala Ser Asp Pro Leu Val Met Tyr Gly Asn370 375 380Val Tyr Asp Ile Thr Asn Wing Be Asp Pro Read Val Met Tyr Gly Asn370 375 380

Gln Asp Glu Glu Ala Asp Arg Ser Leu Ser His Glu Ser Lys Leu Asp385 390 395 400Gln Asp Glu Glu Wing Asp Arg Be Read His Be Glu Be Lys Read Asp385 390 395 400

Phe Gly Asp Pro Phe Arg Lys His Lys Met Ile Cys Arg Tyr His Gln 405 410 415Phe Gly Asp Pro Phe Arg Lys His Lys Met Ile Cys Arg Tyr His Gln 405 410 415

Lys Ala Pro Ile Pro Leu Asn vai Leu Thr Thr Val Pro Leu Phe PheLys Wing Pro Ile Pro Read Asn Goes Read Thr Thr Val Pro Read Phe Phe

420 425 430420 425 430

Ala Ile Gly Phe Leu vai Gly Tyr Ile Leu Tyr Gly Ala Ala Met His435 440 445Ile Wing Gly Phe Leu Goes Gly Tyr Ile Leu Tyr Gly Wing Met Wing His435 440 445

Ile vai Lys Val Glu Asp Asp Phe His Glu Met Gln Glu Leu Lys Val450 455 460Ile Goes Lys Val Glu Asp Asp Phe His Glu Met Gln Glu Read Lys Val450 455 460

Arg Ala Glu Ala Ala Asp Val Ala Lys Ser Gln Phe Leu Ala Thr Val465 470 475 480Arg Wing Glu Wing Wing Asp Val Wing Lys Ser Gln Phe Leu Wing Thr Val465 470 475 480

Ser His Glu Ile Arg Thr Pro Met Asn Gly lie Leu Gly Met Leu Ala 485 490 495Be His Glu Ile Arg Thr Pro Met Asn Gly lie Leu Gly Met Leu Wing 485 490 495

Met Leu Leu Asp Thr Glu Leu Ser Ser Thr Gln Arg Asp Tyr Ala Gln500 505 510Thr Ala Gln vai Cys Gly Lys Ala Leu Ile Ala Leu Ile Asn Glu vaiMet Leu Read Asp Thr Glu Read Be Being Thr Gln Arg Asp Tyr Wing Gln500 505 510Thr Wing Gln Go Cys Gly Lys Wing Leu Ile Wing Leu Ile Asn Glu Go

515 520 525515 520 525

Leu Asp Arg Ala Lys lie Glu Ala Gly Lys Leu Glu Leu Glu Ser Val530 535 540Leu Asp Arg Wing Lys lie Glu Wing Gly Lys Leu Glu Leu Glu Ser Val530 535 540

Pro Phe Asp Ile Arg Ser Ile Leu Asp Asp vai Leu Ser Leu Phe Ser545 550 555 560Pro Phe Asp Ile Arg Be Ile Read Asp Asp Will Read Be Read Phe Ser545 550 555 560

Glu Glu Ser Arg Asn Lys Gly Ile Glu Leu Ala Val Phe Val Ser AspGlu Glu Ser Arg Asn Lys Gly Ile Glu Leu Wing Val Phe Val Ser Asp

565 570 575565 570 575

Lys Val Pro Glu Ile Val Lys Gly Asp Ser Gly Arg Phe Arg Gln Ile 580 585 590Lys Val Pro Glu Ile Val Lys Gly Asp Be Gly Arg Phe Arg Gln Ile 580 585 590

Ile lie Asn Leu Val Gly Asn Ser vai Lys Phe Thr Glu Lys Gly HisIle lie Asn Leu Val Gly Asn Ser will Lys Phe Thr Glu Lys Gly His

595 600 605595 600 605

Ile Phe Val Lys vai His Leu Ala Glu Gln Ser Lys Asp Glu Ser Glu610 615 620Ile Phe Val Lys Goes His Leu Wing Glu Gln Ser Lys Asp Glu Ser Glu610 615 620

Pro Lys Asn Ala Leu Asn Gly Gly Val Ser Glu Glu Met Ile Val vai625 630 635 640Pro Lys Asn Wing Read Asn Gly Gly Val Ser Glu Glu Met Ile Val vai625 630 635 640

Ser Lys Gln Ser Ser Tyr Asn Thr Leu Ser Gly Tyr Glu Ala Ala AspSer Lys Gln Ser Ser Tyr Asn Thr Read Ser Ser Gly Tyr Glu Ala Wing Asp

645 650 655645 650 655

Gly Arg Asn Ser Trp Asp Ser Phe Lys His Leu vai Ser Glu Glu Gln 660 665 670Gly Arg Asn Be Trp Asp Be Phe Lys His Leu Will Be Glu Glu Gln 660 665 670

Ser Leu ser Glu Phe Asp Ile ser Ser Asn vai Arg Leu Met Val serBe Leu be Glu Phe Asp Ile be Ser Asn will Arg Leu Met Val be

675 680 685675 680 685

Ile Glu Asp Thr Gly Ile Gly Ile Pro Leu Val Ala Gln Gly Arg Val690 695 700Ile Glu Asp Thr Gly Ile Gly Ile Pro Read Val Wing Gln Gly Arg Val690 695 700

Phe Met Pro Phe Met Gln Ala Asp Ser ser Thr Ser Arg Asn Tyr Gly705 710 715 720Phe Met Pro Phe Met Gln Wing Asp Be Be Thr Be Arg Asn Tyr Gly705 710 715 720

Gly Thr Gly lie Gly Leu Ser Ile Ser Lys Cys Leu Val Glu Leu MetGly Thr Gly lie Gly Leu Be Ile Ser Lys Cys Leu Val Glu Leu Met

725 730 735725 730 735

Arg Gly Gln Ile Asn Phe Ile Ser Arg Pro His Ile Gly Ser Thr Phe 740 745 750Arg Gly Gln Ile Asn Phe Ile Be Arg Pro His Ile Gly Be Thr Phe 740 745 750

Trp Phe Thr Ala vai Leu Glu Lys Cys Asp Lys Cys Ser Ala Ile Asn755 760 765His Met Lys Lys Pro Asn Val Glu His Leu Pro Ser Thr Phe Lys Gly770 775 780Trp Phe Thr Wing Go Read Glu Lys Cys Asp Lys Cys Be Wing Ile Asn755 760 765His Met Lys Lys Pro Asn Val Glu His Leu Pro Be Thr Phe Lys Gly770 775 780

Met Lys Ala lie Val vai Asp Ala Lys Pro vai Arg Ala Ala Val Thr785 790 795 800Met Lys Ala lie Val goes Asp Alys Lys Pro goes Arg Ala Val Wing Thr785 790 795 800

Arg Tyr His Met Lys Arg Leu Gly Ile Asn vai Asp vai Val Thr ser805 810 815Arg Tyr His Met Lys Arg Leu Gly Ile Asn Goes Asp Goes Val Thr be805 810 815

Leu Lys Thr Ala Val vai Ala Ala Ala Ala Phe Glu Arg Asn Gly Ser820 825 830Leu Lys Thr Wing Val goes Wing Wing Wing Wing Phe Glu Arg Wing Asn Gly Ser820 825 830

Pro Leu Pro Thr Lys Pro Gln Leu Asp Met Ile Leu Val Glu Lys Asp835 840 845Pro Read Pro Thr Lys Pro Gln Read Asp Met Ile Read Val Glu Lys Asp835 840 845

Ser Trp Ile Ser Thr Glu Asp Asn Asp Ser Glu Ile Arg Leu Leu Asn850 855 860Be Trp Ile Be Thr Glu Asp Asn Asp Be Glu Ile Arg Leu Read Asn850 855 860

Ser Arg Thr Asn Gly Asn vai His His Lys Ser Pro Lys Leu Ala Leu865 870 875 880Be Arg Thr Asn Gly Asn Goes His His Lys Be Pro Lys Leu Wing Leu865 870 875 880

Phe Ala Thr Asn Ile Thr Asn Ser Glu Phe Asp Arg Ala Lys Ser Ala885 890 895Phe Ala Thr Asn Ile Thr Asn Be Glu Phe Asp Arg Ala Lys Be Ala885 890 895

Gly Phe Ala Asp Thr Val Ile Met Lys Pro Leu Arg Ala Ser Met Ile900 905 910Gly Phe Wing Asp Thr Val Ile Met Lys Pro Read Arg Wing Be Met Ile900 905 910

Gly Ala Cys Leu Gln Gln vai Leu Glu Leu Arg Lys Thr Arg Gln Gln915 920 925Gly Wing Cys Read Gln Gln Go Read Glu Read Le Arg Lys Thr Arg Gln915 920 925

His Pro Glu Gly Ser Ser Pro Ala Thr Leu Lys Ser Leu Leu Thr Gly930 935 940His Pro Glu Gly Be Be Pro Wing Thr Read Lys Be Read Leu Thr Gly930 935 940

Lys Lys Ile Leu Val Val Asp Asp Asn Ile Val Asn Arg Arg Val Ala945 950 955 960Lys Lys Ile Leu Val Val Asp Asp Asn Ile Val Asn Arg Arg Val Ala945 950 955 960

Ala Gly Ala Leu Lys Lys Phe Gly Ala Glu Val Val Cys Ala Glu Ser965 970 975Wing Gly Wing Read Lys Lys Phe Gly Glu Wing Val Val Cys Glu Wing Ser965 970 975

Gly Gln Val Ala Leu Gly Leu Leu Gln Ile Pro His Thr Phe Asp Ala980 985 990Gly Gln Val Wing Read Gly Leu Read Gln Ile Pro His Thr Phe Asp Wing 980 985 990

Cys Phe Met Asp Ile Gln Met Pro Gln Met Asp Gly Phe Glu Ala Thr995 1000 1005Cys Phe Met Asp Ile Gln Met Pro Gln Met Asp Gly Phe Glu Wing Thr995 1000 1005

Arg Gln Ile Arg Met Met Glu Lys Glu Ala Lys Glu Lys Thr Asn1010 1015 1020Leu Glu Trp His Leu Pro Ile Leu Ala Met Thr Ala Asp vai IleArg Gln Ile Arg Met Met Glu Lys Glu Wing Lys Glu Lys Thr Asn1010 1015 1020Leu Glu Trp His Leu Pro Ile Leu Wing Met Thr Wing Asp Go Ile

1025 1030 10351025 1030 1035

His Ala Thr Tyr Glu Glu Cys Leu Lys Ser Gly Met Asp Gly Tyr1040 1045 1050His Wing Thr Tyr Glu Glu Cys Read Lys Ser Gly Met Asp Gly Tyr1040 1045 1050

Val Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr Lys Ser Val Ala1055 1060 1065Val Ser Lys Pro Phe Glu Glu Glu Asn Leu Tyr Lys Ser Val Ala1055 1060 1065

Lys Ser Phe Lys Pro Asn Pro Ile Ser Pro ser SerLys Ser Phe Lys Pro Asn Pro

1070 1075 10801070 1075 1080

<210> 18<211> 1013<212> PRT<213> Oryza<220><210> 18 <211> 1013 <212> PRT <213> Oryza <220>

<221> Mutation<222> (266)..(266)<221> Mutation <222> (266) .. (266)

<223> X=Xaa (any amino acid other than Leu)<220><223> X = Xaa (any amino acid other than Leu) <220>

<221> misc_feature<222> (266)..(266)<223> Xaa can be any naturalIy occurring amino acid<400> 18<221> misc_feature <222> (266) .. (266) <223> Xaa can be any naturalIy occurring amino acid <400> 18

Met Gly Val Gly Gly Gly Gly Gly Gly Gly Gly Gly Glu Ala Ala Ala1 5 10 15Met Gly Val Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly

Ala vai Ala vai Glu Gly Asp Glu Ala Gly Lys Gly Arg Arg Trp Trp20 25 30Wing Go Wing Go Glu Gly Asp Glu Wing Gly Lys Gly Arg Arg Trp Trp20 25 30

Arg vai Lys vai Lys Leu ser Thr vai Ala vai vai Ala Trp Val LeuArg will Lys will Lys Leu be Thr will Wing will go Wing Trp Val Leu

35 40 4535 40 45

Ala Ser Ala Ala Leu Trp Ala Gly Leu His Trp Arg Phe Arg Arg Ala50 55 60Wing Be Wing Wing Read Trp Wing Gly Read His Trp Arg Phe Arg Arg Wing50 55 60

Ala Leu His Lys Ala Glu Glu Ala Leu Val Cys Met Cys Glu Glu Arg65 70 75 80Wing Read His Lys Wing Glu Glu Wing Read Leu Val Cys Met Cys Glu Glu Arg65 70 75 80

Ala Arg Met Leu Gln Asp Gln Phe Ala vai Ser Val Asn His Val His85 90 95Wing Arg Met Read Gln Asp Gln Phe Wing Will Be Val Asn His Val His85 90 95

Ala Leu Ala lie Leu vai Ala Thr Phe His Tyr Asp Lys His Pro ProAla Leu Ala lie Leu goes Ala Thr Phe His Tyr Asp Lys His Pro

100 105 110100 105 110

Ala Leu Asp Gln Asp Thr Phe Ala Val Tyr Ala Ala Arg Thr Ser Phe115 120 125Wing Read Asp Gln Asp Thr Phe Wing Val Tyr Wing Wing Arg Thr Be Phe115 120 125

Glu Arg Pro Leu Leu Ser Gly Val Ala Tyr Ala Gln Arg Val Val HisGlu Arg Pro Leu Read Ser Gly Val Wing Tyr Wing Gln Arg Val Val His

130 135 140130 135 140

Ala Asp Arg Glu Ser Phe Glu Arg Gln Gln Gly Trp Ile Ile Lys Thr145 150 155 160Wing Asp Arg Glu Be Phe Glu Arg Gln Gln Gly Trp Ile Ile Lys Thr145 150 155 160

Met Lys His Glu Pro ser Pro Ala Gln Asp Glu Tyr Ala Pro Val IleMet Lys His Glu Pro Be Pro Wing Gln Asp Glu Tyr Wing Pro Val Ile

165 170 175165 170 175

Tyr Ser Gln Glu Thr Ile Ser Tyr Ile Glu Gly Leu Asp Val Met SerTyr Ser Gln Glu Thr Ile Ser Tyr Ser Glu Gly Leu Asp Val Met Ser

180 185 190180 185 190

Gly Glu Glu Asp Arg Glu Asn Ile Leu Arg Ala Arg Ala Thr Gly Lys195 200 205Gly Glu Glu Asp Arg Glu Asn Ile Leu Arg Wing Arg Wing Wing Thr Gly Lys195 200 205

Ala Val Leu Thr Arg Pro Phe Arg Leu Met Ser Asn His Leu Gly ValWing Val Leu Thr Arg Pro Phe Arg Leu Met Ser Asn His Leu Gly Val

210 215 220210 215 220

vai Leu Thr Phe Pro Val Tyr Leu Val Asp Leu Pro Asn Asp Thr Ala225 230 235 240go Read Thr Phe Pro Val Tyr Read Val Asp Read Le Pro Asn Thr Ala225 230 235 240

Val Glu Asp Arg Val Ala Ala Thr Ala Gly Tyr Leu Gly Gly Ala PheVal Glu Asp Arg Val Wing Ward Wing Thr Wing Gly Tyr Leu Gly Gly Wing Phe

245 250 255245 250 255

Asp Val Glu Ser Leu vai Glu Asn Leu Xaa Arg Gln Leu Ala Gly AsnAsp Val Glu Being Leu Goes Glu Asn Leu Xaa Arg Gln Leu Wing Gly Asn

260 265 270260 265 270

Gln Glu Leu vai vai Asn Val Tyr Asp Val Thr Asn His Ser Asn Pro275 280 285Gln Glu Leu Goes Go Asn Val Tyr Asp Val Thr Asn His Ser Asn Pro275 280 285

Leu vai Met Tyr Gly Ser Glu Val Pro Leu Gly Ile Pro Ser Pro SerLeu Goes Met Tyr Gly To Be Glu Val Pro Leu Gly Ile To Be Pro Pro

290 295 300290 295 300

His Thr Tyr Thr Leu Asp Phe Gly Asp Pro Leu Arg Lys His Gln Met305 310 315 320His Thr Tyr Thr Read Asp Phe Gly Asp Pro Read Arg Lys His Gln Met305 310 315 320

Val Cys Arg Tyr Arg Asn Lys Leu His vai Ser Trp ser Ala Ile ThrVal Cys Arg Tyr Arg Asn Lys Leu His Will Be Trp Be Wing Ile Thr

325 330 335325 330 335

Thr Pro Ser Gly Val Phe vai Ile Cys Met Leu Val Gly Tyr Ile Ile340 345 350Thr Pro Be Gly Val Phe Goes Ile Cys Met Read Val Gly Tyr Ile Ile340 345 350

Tyr Ala Ala Trp Ser Arg Tyr Asp Asn vai Lys Glu Asp Cys Arg LysTyr Ala Wing Trp Being Arg Tyr Asp Asn Goes Lys Glu Asp Cys Arg Lys

355 360 365355 360 365

Met Glu Ala Leu Lys Lys Arg Ala Glu Ala Ala Asp Ile Ala Lys Ser370 375 380Met Glu Wing Leu Lys Lys Arg Wing Glu Wing Wing Asp Ile Wing Lys Ser370 375 380

Gln Phe Leu Ala Thr Val Ser His Glu Ile Arg Thr Pro Met Asn Gly385 390 395 400Gln Phe Read Wing Thr Val Be His Glu Ile Arg Thr Pro Met Asn Gly385 390 395 400

vai Leu Gly Met Leu Asp Met Leu Leu Asp Thr Glu Leu Lys Ser Thr405 410 415go Read Gly Met Read Asp Met Read Leu Read Asp Thr Glu Read Le Lys Ser Thr405 410 415

Gln Arg Asp Tyr Ala Gln Thr Ala Gln Val Cys Gly Lys Ala Leu Ile420 425 430Gln Arg Asp Tyr Wing Gln Thr Wing Gln Val Cys Gly Lys Wing Leu Ile420 425 430

Ser Leu Ile Asn Glu vai Leu Asp Arg Ala Lys Ile Glu Ala Gly LysSer Leu Ile Asn Glu will Leu Asp Arg Wing Lys Ile Glu Wing Gly Lys

435 440 445435 440 445

Ile Asp Leu Glu Ser Val Pro Phe Asp Leu Arg Ser Ile Leu Asp Asp450 455 460Ile Asp Leu Glu Be Val Pro Phe Asp Leu Arg Be Ile Leu Asp Asp450 455 460

vai Ile Ser Leu Phe Ser Ser Lys Ser Arg Glu Lys Gly Ile Glu Leu465 470 475 480will Ile Be Leu Phe Be Be Lys Be Arg Glu Lys Gly Ile Glu Leu465 470 475 480

Ala Val Tyr Val Ser Glu Arg Val Pro Glu Ile Leu Leu Gly Asp Pro485 490 495Val Tyr Wing Val Ser Glu Arg Val Pro Glu Ile Leu Read Gly Asp Pro485 490 495

Gly Arg Phe Arg Gln Ile lie Thr Asn Leu Val Gly Asn ser Ile Lys500 505 510Gly Arg Phe Arg Gln Ile Lie Thr Asn Read Val Gly Asn Be Ile Lys500 505 510

Ile Thr Ile Phe Thr Leu Ser Gln Phe Thr Glu Arg Gly His Ile PheIle Thr Ile Phe Thr Read Be Gln Phe Thr Glu Arg Gly His Ile Phe

515 520 525515 520 525

Val Gln Val His Leu Ala Asp His Ser Asn Leu Ala Thr Glu Ala Lys530 535 540Val Gln Val His Leu Wing Asp His Ser Asn Leu Wing Wing Thr Glu Wing Lys530 535 540

Ile Glu Pro vai vai Asn Gly Met Asn Gly His Lys Asp Glu Ala Ile545 550 555 560Ile Glu Pro Goes Go Asn Gly Met Asn Gly His Lys Asp Glu Wing Ile545 550 555 560

Ala Ile Pro Thr Ser Gly Ser His Asn Thr Leu Ser Gly Phe Glu Ala565 570 575Wing Ile Pro Thr Be Gly Be His Asn Thr Read Be Gly Phe Glu Ala565 570 575

Ala Asp Ser Arg Asn Asn Trp Glu Asn Phe Lys Leu Leu Leu Ser Tyr580 585 590Wing Asp Ser Arg Asn Asn Trp Glu Asn Phe Lys Leu Leu Leu Ser Tyr580 585 590

Glu Lys Asn Glu Met Pro Tyr Glu Ser Asp Ser Asp Lys vai Thr Leu595 600 605Glu Lys Asn Glu Met Pro Tyr Glu Be Asp Be Asp Lys Go Thr Leu595 600 605

VaT val ser vai Glu Asp Thr Gly lie Gly Ile Pro Leu His Ala GlnVaT val be go Glu Asp Thr Gly lie Gly Ile Pro Read His Wing Gln

610 615 620610 615 620

Gly Arg Val Phe Thr Pro Phe Met Gln Ala Asp Ser Ser Thr Ser Arg625 630 635 640Gly Arg Val Phe Thr Pro Phe Met Gln Wing Asp Be Be Thr Be Arg625 630 635 640

Asn Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile Ser Lys Cys Leu ValAsn Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile Ser Lys Cys Leu Val

645 650 655645 650 655

Glu Ile Met Gly Gly Gln Ile Asn Phe vai Ser Arg Pro Leu vai GlyGlu Ile Met Gly Gly Gln Ile Asn Phe Will Be Arg Pro Leu Go Gly

660 665 670660 665 670

10 Ser Thr Phe Thr Phe Thr Ala Val Leu Arg Arg Cys Asp Lys Asn Ala675 680 68510 Ser Thr Phe Thr Phe Thr Wing Val Leu Arg Arg Cys Asp Lys Asn Ala675 680 685

Ile Ser Asp Ser Lys Thr Val Ala Leu His Pro Leu Pro Ser Ser PheIle Be Asp Be Lys Thr Val Wing Read His Pro Read Pro Be Ser Phe

690 695 700690 695 700

Lys Gly Leu Ser Ala Leu Leu Val Asp Lys Arg Pro vai Arg Ala Thr705 710 715 720Lys Gly Leu Be Wing Leu Leu Val Asp Lys Arg Pro Goes Arg Wing Thr705 710 715 720

vai Thr Lys Tyr His Leu Gln Arg Leu Gly Ile Thr ser Glu vai vaiGo Thr Lys Tyr His Glu Arg Leu Gly Gle Ile Thr Be Glu Go

725 730 735725 730 735

Gly Thr Ile Asp Pro Thr Phe Gly Val Leu Ser Gly Arg Asn Gly ser740 745 750Gly Thr Ile Asp Pro Thr Phe Gly Val Leu Ser Gly Arg Asn Gly ser740 745 750

Ser Leu Thr Ser Ile Gly Lys Lys Gln Pro Cys Met Leu Leu Ile Glu755 760 765Be Leu Thr Be Ile Gly Lys Lys Gln Pro Cys Met Read Leu Ile Glu755 760 765

ser Asp Ser Trp Gly Pro Gln Met Asp vai Ser Leu His Ala Arg LeuBe Asp Be Trp Gly Pro Gln Met Asp Will Be Read His Wing Arg Read

770 775 780770 775 780

Gln Glu Met Lys Gln Ser Asp Arg Ile His vai Leu Pro Lys Val Phe785 790 795 800Gln Glu Met Lys Gln Being Asp Arg Ile His Going To Read Pro Lys Val Phe785 790 795 800

Leu Leu Ser Ala Ala Glu Ser Asp Lys Val Lys Lys Ile His Ala vaiLeu Leu Be Wing Wing Glu Wing Asp Lys Val Lys Lys Ile His Wing Goes

805 810 815805 810 815

Asp Ser vai Ile Pro Lys Pro Leu Lys Ala Ser Ala Leu Ala Ala Cys820 825 830Asp Will Go Ile Pro Lys Pro Read Lys Wing Be Wing Read Wing Wing Cys820 825 830

30 Leu Phe Gln Ala Leu Gly Ile Thr Gln Pro ser His Glu Lys Arg Asp835 840 84530 Leu Phe Gln Wing Leu Gly Ile Thr Gln Pro Be His Glu Lys Arg Asp835 840 845

Asp Ser Gly Ser Leu His Gly Arg Asp Gly Ser Gly ser Leu His Gly850 855 860Asp Be Gly Be Read His Gly Arg Asp Gly Be Read His Gly Arg 855 860

Leu Leu Leu Gly Lys Asn lie Leu vai vai Asp Asp Asn Lys Val Asn865 870 875 880Leu Leu Leu Gly Lys Asn lie Leu will go Asp Asp Asn Lys Val Asn865 870 875 880

Leu Arg Val Ala Ala Gly Thr Leu Lys Lys Tyr Gly Ala Lys Val Glu 885 890 895Leu Arg Val Wing Wing Gly Thr Read Leys Lys Lys Tyr Gly Wing Lys Val Glu 885 890 895

Cys Val Glu ser Gly Lys Asp Ala Leu Ser Leu Leu Gln Val Pro HisCys Val Glu Be Gly Lys Asp Wing Read Le Be Read Leu Gln Val Pro His

900 905 910900 905 910

Lys Phe Asp Leu Cys Leu Met Asp Ile Gln Met Pro Glu Met Asp Gly915 920 925Lys Phe Asp Read Cys Read Met Asp Ile Gln Met Pro Glu Met Asp Gly915 920 925

Phe Glu Ala Thr Arg Gln Ile Arg Ala Met Glu Gly Lys Ala Asn Glu930 935 940Phe Glu Wing Thr Arg Gln Ile Arg Wing Met Glu Gly Lys Wing Asn Glu930 935 940

Gln Ala Asp Asp ser Glu Ser Gly Ser Glu Ile Ala Ala Lys Thr Ala945 950 955 960Gln Wing Asp Asp Be Glu Be Gly Be Glu Ile Wing Wing Lys Thr Wing 945 950 955 960

Lys Trp His Leu Pro Ile Leu Ala Met Thr Ala Asp Val Ile Gln Ala 965 970 975Lys Trp His Leu Pro Ile Leu Wing Met Thr Wing Asp Val Ile Gln Wing 965 970 975

Thr His Glu Glu Cys Thr Lys Cys Gly Met Asp Gly Tyr vai Ser LysThr His Glu Glu Cys Thr Lys Cys Gly Met Asp Gly Tyr Will Be Lys

980 985 990980 985 990

Pro Phe Glu Glu Lys Gln Leu Phe Gln Ala vai Gln Lys Phe Leu Gly995 1000 1005Pro Phe Glu Glu Lys Gln Leu Phe Gln Wing Goes Gln Lys Phe Leu Gly995 1000 1005

Pro Cys vai Ser Ser1010Pro Cys Will Be Ser1010

<210> 19<211> 995<212> PRT<213> Zea mays<220><210> 19 <211> 995 <212> PRT <213> Zea mays <220>

<221> Mutation<222> (267)..(267)<221> Mutation <222> (267) .. (267)

<223> X= Xaa (any amino acid other than Leu)<220><223> X = Xaa (any amino acid other than Leu) <220>

<221> misc_feature<222> (267)..(267)<223> Xaa can be any naturally occurring amino acid<400> 19<221> misc_feature <222> (267) .. (267) <223> Xaa can be any naturally occurring amino acid <400> 19

Met Gly Val Gly Gly Gly Gly Gly Gly Glu Ala Ala Ala Val Ser Ala1 5 10 15Met Gly Gly Gly Gly Gly Gly Gly Gly Glu Wing Wing Wing Val Ser Wing1 5 10 15

Pro Ala Pro Ala Glu Glu Ala Gly Lys Asp Ala Glu Asp Gly Gly Gly20 25 30Pro Wing Pro Wing Glu Glu Wing Gly Lys Asp Wing Glu Asp Gly Gly Gly20 25 30

Trp Thr Leu Lys Ala Lys Leu Ile Ala Val Ala Val Leu Val Trp ValTrp Thr Leu Lys Wing Lys Leu Ile Val Wing Val Wing Val Leu Val Trp Val

35 40 4535 40 45

Leu Gly Ala Leu Ala Leu Gly Val Phe Leu His Ser Tyr Phe Arg His50 55 60Leu Gly Wing Leu Wing Leu Gly Val Phe Leu His Ser Tyr Phe Arg His50 55 60

Ala Ala Leu Arg Lys Ala Glu Glu Gly Leu Val Ser Met Cys Glu Glu65 70 75 80Wing Wing Leu Arg Lys Wing Glu Glu Gly Leu Val Ser Met Cys Glu Glu65 70 75 80

Arg Ala Arg Met Leu Gln Asp Gln Phe Ala vai Ser Val Asn His Val85 90 95Arg Wing Arg Met Leu Gln Asp Gln Phe Wing Will Be Val Asn His Val85 90 95

His Ala Leu Ala Ile Leu Val Ala Thr Phe His Tyr Glu Lys Arg Pro100 105 110His Wing Leu Wing Ile Leu Val Val Thr Phe His Tyr Glu Lys Arg Pro100 105 110

Pro Ala Leu Asp Gln Asn Thr Phe Ala Asp Tyr Thr Ala Arg Thr SerPro Wing Read Asp Gln Asn Thr Phe Wing Asp Tyr Thr Wing Arg Thr

115 120 125115 120 125

Phe Glu Arg Pro Leu Leu ser Gly vai Ala Tyr Ala Gln Arg Val Val130 135 140Phe Glu Arg Pro Leu Read Gly Go Gly Wing Tyr Wing Gln Arg Val Val130 135 140

His Gly Asp Arg Glu Ser Phe Glu Arg Gln Gln Gly Trp Ile Ile Lys145 150 155 160His Gly Asp Arg Glu Being Phe Glu Arg Gln Gln Gly Trp Ile Ile Lys145 150 155 160

Thr Met Lys His Glu Pro Ser Pro Val Gln Asp Glu Tyr Ala Pro Val165 170 175Thr Met Lys His Glu Pro Pro Val Gln Asp Glu Tyr Wing Pro Val165 170 175

vai Tyr Ser Gln Glu Thr vai Ser Tyr Ile Glu Gly Leu Asp Met Met180 185 190will Tyr Be Gln Glu Thr will be Tyr Ile Glu Gly Leu Asp Met Met180 185 190

Ser Gly Glu Glu Asp Arg Glu Asn Ile Leu Arg Ser Arg Ala Ser GlyBe Gly Glu Glu Asp Arg Glu Asn Ile Leu Arg Be Arg Wing Be Gly

195 200 205195 200 205

Lys Ala Val Leu Thr Arg Pro Phe Arg Leu Met Ser Asn His Leu Gly210 215 220Lys Wing Val Leu Thr Thr Pro Phe Arg Leu Met Ser Asn His Leu Gly210 215 220

vai vai Leu Thr Phe Pro vai Tyr His Val Asp Leu Ser Ser Asp Ala225 230 235 240Lys Glu Glu Asp Arg Val Ala Ala Thr Ala Gly Tyr Leu Gly Gly Ser245 250 255Go Go Leu Thr Phe Pro Go Tyr His Val Asp Leu Be Ser Asp Ala225 230 235 240Lys Glu Glu Asp Arg Val Wing Ala Thr Wing Gly Tyr Leu Gly Gly Ser245 250 255

Phe Asp Val Glu Ser Leu vai Glu Asn Leu Xaa Arg Gln Leu Ala Gly260 265 270Phe Asp Val Glu Being Leu Goes Glu Asn Leu Xaa Arg Gln Leu Wing Gly260 265 270

Asn Gln Glu Leu Val Val Asn vai Tyr Asp Val Thr Asn Ser Ser Asn275 280 285Asn Gln Glu Leu Val Val Asn will Tyr Asp Val Thr Asn Be Ser Asn275 280 285

Pro Leu Val Met Tyr Gly Ser Glu vai Ser Leu Gly Asn Pro Ser ProPro Read Val Met Tyr Gly Be Glu Will Be Read Gly Asn Pro Be Pro

290 295 300290 295 300

Ser His Ile Cys Met Leu Asp Phe Gly Asp Pro Phe Arg Lys His His305 310 315 320Be His Ile Cys Met Read Asp Phe Gly Asp Pro Phe Arg Lys His His305 310 315 320

Met Val Cys Arg Tyr Arg Asn Lys Pro Gln Leu Pro Trp Ser Ala Ile325 330 335Met Val Cys Arg Tyr Arg Asn Lys Pro Gln Read Pro Trp Be Wing Ile325 330 335

Ser Ser Ser Ser Gly Val Phe Val Ile Cys Met Leu Val Gly Tyr Ile340 345 350Being Being Being Being Gly Val Phe Val Ile Cys Met Leu Val Gly Tyr Ile340 345 350

Val Gly Ala Ala Trp Ser Arg Tyr Asp Asn Val Lys Glu Asp Cys Arg355 360 365Val Gly Wing Trp Wing Be Arg Tyr Asp Asn Val Lys Glu Asp Cys Arg355 360 365

Lys Met Glu Glu Leu Lys Lys Gln Ala Glu Ala Ala Asp Val Ala Lys370 375 380Lys Met Glu Glu Leu Lys Lys Gln Wing Glu Wing Wing Asp Val Wing Lys370 375 380

Ser Gln Phe Leu Ala Thr Val Ser His Glu Ile Arg Thr Pro Met Asn385 390 395 400Be Gln Phe Leu Thr Wing Val Be His Glu Ile Arg Thr Pro Met Asn385 390 395 400

Gly Val Leu Gly Met Leu Asp Met Leu Leu Asp Thr Asp Leu Thr Ser405 410 415Gly Val Leu Gly Met Leu Asp Met Leu Read Asp Thr Asp Leu Thr Ser405 410 415

Thr Gln Arg Asp Phe Ala Gln Thr Ala Gln Val Cys Gly Lys Ala Leu420 425 430Thr Gln Arg Asp Phe Wing Gln Thr Wing Gln Val Cys Gly Lys Wing Leu420 425 430

Ile Ser Leu Ile Asn Glu vai Leu Asp Arg Ala Lys Ile Glu Ala Gly435 440 445Ile Ser Leu Ile Asn Glu will Read Asp Arg Wing Lys Ile Glu Wing Gly435 440 445

Lys Leu Asp Leu Glu Ser Val Pro Phe Asp Leu Arg Ser Ile Leu Asp450 455 460Lys Leu Asp Leu Glu Be Val Pro Phe Asp Leu Arg Be Ile Leu Asp450 455 460

Asp Val Ile Ser Leu Phe Ser ser Lys ser Arg Glu Lys Gly Ile Glu465 470 475 480Asp Val Ile Be Leu Phe Be Be Lys Be Arg Glu Lys Gly Ile Glu465 470 475 480

Leu Ala Val Tyr vai Ser Glu Arg Val Pro Glu Leu Leu Leu Gly Asp485 490 495Pro Gly Arg Phe Arg Gln Ile Ile Thr Asn Leu vai Gly Asn Ser IleLeu Wing Val Tyr Will Be Glu Arg Val Pro Glu Leu Leu Leu Gly Asp485 490 495Pro Gly Arg Phe Arg Gln Ile Ile Thr Asn Leu Goes Gly Asn Ser Ile

500 505 510500 505 510

Lys Phe Thr Glu Arg Gly His Ile Phe Val Gln Val His Leu Ala Asp515 520 525Lys Phe Thr Glu Arg Gly His Ile Phe Val Gln Val His Leu Wing Asp515 520 525

His Ser Asn Leu Ala Thr Glu Ser Lys Val Glu Ser Val Ala Asn Gly530 535 540His Ser Asn Leu Wing Thr Glu Ser Lys Val Glu Ser Val Wing Asn Gly530 535 540

Met Asn Gly His Lys Asp Glu Lys Thr Ala vai Ala Thr Ser Val Ser545 550 555 560Met Asn Gly His Lys Asp Glu Lys Thr Wing Go Wing Thr Be Val Ser545 550 555 560

Leu Asn Thr Leu Ser Gly Phe Glu Ala Ala Asp Ser Arg Asn Ser Trp565 570 575Leu Asn Thr Leu Be Gly Phe Glu Wing Wing Asp Be Arg Asn Be Trp565 570 575

Glu Asn Phe Lys Leu Leu Leu Ser Tyr Glu Lys Asn Glu Met Pro TyrGlu Asn Phe Lys Leu Leu Leu Ser Tyr Glu Lys Asn Glu Met Pro Tyr

580 585 590580 585 590

Glu Ser Val Ser Asp Lys vai Thr Leu Val Val Ser Val Glu Asp Thr595 600 605Glu Ser Val Ser Asp Lys Go Thr Leu Val Val Ser Val Glu Asp Thr595 600 605

Gly Ile Gly Ile Pro Leu Asp Ala Gln Ala Lys vai Phe Thr Pro Phe610 615 620Gly Ile Gly Ile Pro Read Asp Wing Gln Wing Lys Go Phe Thr Pro Phe610 615 620

Met Gln Ala Asp Ser Ser Thr Ser Arg Thr Tyr Gly Gly Thr Gly Ile625 630 635 640Met Gln Wing Asp Be Be Thr Be Arg Be Tyr Gly Gly Thr Gly Ile625 630 635 640

Gly Leu Ser Ile Ser Lys Cys Leu vai Glu Leu Met Gly Gly Gln Ile645 650 655Gly Leu Being Ile Being Lys Cys Leu Goes Glu Leu Met Gly Gly Gln Ile645 650 655

Asn Phe vai Ser Arg Pro His vai Gly Ser Thr Phe Thr Phe Thr AlaAsn Phe Will Be Arg Pro His Will Gly Be Thr Phe Thr Phe Thr Wing

660 665 670660 665 670

Ala Leu Gln Arg Cys Asp Arg Ser Ala Ile Gly Asp Ser Lys Pro Val675 680 685Wing Read Gln Arg Cys Asp Arg Be Wing Ile Gly Asp Be Lys Pro Val675 680 685

Met Leu His Pro Leu Pro Ser Ser Phe Lys Gly Leu Ser Ala Leu Leu690 695 700Met Leu His Pro Leu Pro To Be Phe Lys Gly Leu To Be Wing Leu Leu690 695 700

Val Asp Arg Arg Pro Val Arg Ala Thr vai Thr Lys Tyr His Leu Gln705 710 715 720Val Asp Arg Arg Pro Val Arg Wing Thr Thr Go Lys Tyr His Leu Gln705 710 715 720

Arg Leu Gly Ile Ala Cys Asp vai vai Ala Thr Ile Glu Leu Ala Leu725 730 735Arg Leu Gly Ile Wing Cys Asp Goes Go Wing Thr Ile Glu Leu Wing Leu725 730 735

Gly Val Leu Ser Gly Arg Asn Gly Ser Ser Leu Thr Ser Thr Lys Gln740 745 750Pro Cys Met Leu Leu Ile Glu Ser Asp Ser Trp Gly Phe Lys Ile AspGly Val Leu Be Gly Arg Asn Gly Be Be Leu Thr Be Thr Lys Gln740 745 750Pro Cys Met Leu Read Ile Glu Be Asp Be Trp Gly Phe Lys Ile Asp

755 760 765755 760 765

Val Pro Leu Arg Ser Arg Leu Leu Glu Met Lys Gln Asn Gly Pro Pro770 775 780Val Pro Read Arg Be Arg Read Leu Glu Met Lys Gln Asn Gly Pro Pro770 775 780

Gly Leu Pro Lys Thr Ile Leu Leu Ala Ala Ala Glu Ser Gly Lys Leu785 790 795 800Gly Leu Pro Lys Thr Ile Leu Leu Wing Wing Wing Glu Ser Wing Gly Lys Leu785 790 795 800

Lys Ala His Tyr Ala Val Asp Ser Val Ile Thr Lys Pro Leu Lys AlaLys Wing His Tyr Wing Val Asp Ser Val Ile Thr Lys Pro Read Lys Wing

805 810 815805 810 815

Ser Gly Leu Ala Ala Cys Leu Phe Gln Thr Leu Gly Ile Thr Gln Ser820 825 830Ser Gly Leu Wing Cys Wing Phe Phe Gln Thr Gly Ile Thr Gln Ser820 825 830

Ser Asn Glu Arg Arg Asp Asn Ser Gly Ser Leu His Gly Leu Leu LeuBe Asn Glu Arg Arg Asp Asn Be Gly Be Read His Gly Leu Read Leu

835 840 845835 840 845

Gly Lys Asn Ile Leu Val vai Asp Asp Asn Lys Val Asn Leu Arg Val850 855 860Gly Lys Asn Ile Leu Val Goes Asp Asp Asn Lys Val Asn Leu Arg Val850 855 860

Ala Ala Gly Thr Leu Lys Lys Phe Gly Ala Lys vai Glu Cys Val Glu865 870 875 880Wing Wing Gly Thr Read Lys Lys Lys Phe Gly Wing Lys Goes Glu Cys Val Glu865 870 875 880

Ser Gly Lys Asp Ala Leu Ala Ser Leu Gln Val Pro His Lys Phe HisBe Gly Lys Asp Wing Read Wing Wing Be Read Gln Val Pro His Lys Phe His

885 890 895885 890 895

Leu Cys Leu Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala900 905 910Leu Cys Leu Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala900 905 910

Thr Lys Gln lie Arg Ala Met Glu Ala Lys Ala Asn Glu Gln Ala ValThr Lys Gln lie Arg Wing Met Glu Wing Lys Wing Asn Glu Gln Wing Val

915 920 925915 920 925

Ala Cys Asp Asp Ser Asp Thr Asp Gly Ala Thr Arg Ala Ala Arg Trp930 935 940Cys Wing Asp Asp Be Asp Thr Asp Gly Wing Thr Arg Wing Wing Arg Trp930 935 940

His Leu Pro vai Leu Ala Met Thr Ala Asp vai Ile Gln Ala Thr His945 950 955 960His Leu Pro Goes Leu Wing Met Thr Wing Asp Goes Ile Gln Wing Thr His945 950 955 960

Glu Glu Cys Thr Lys Tyr Gly Met Asp Gly Tyr Val Thr Lys Pro PheGlu Glu Cys Thr Lys Tyr Gly Met Asp Gly Tyr Val Thr Lys Pro Phe

965 970 975965 970 975

Glu Glu Lys Gln Leu Phe Gln Ala Leu Gln Lys Phe Leu Asp Pro Gly980 985 990Glu Glu Lys Gln Read Phe Gln Wing Read Le Gln Lys Phe Read Asp Pro Gly980 985 990

Met Ser Ser995<210> 20Met Ser Ser995 <210> 20

<211> 981<211> 981

<212> PRT<212> PRT

<213> Cucurbita maxima<213> Cucurbita maxima

<220><220>

<221> Mutation<221> Mutation

<222> (261)..(261)<222> (261) .. (261)

<223> X= Xaa (any amino acid other than Leu)<223> X = Xaa (any amino acid other than Leu)

<220><220>

<221> misc_feature<221> misc_feature

<222> (261)..(261)<222> (261) .. (261)

<223> Xaa can be any naturally occurring amino acid<223> Xaa can be any naturally occurring amino acid

<400> 20<400> 20

Met Gln Val Ser Asp Asn Ser Val Gly Leu Lys Trp Asn Glu Gln Met1 5 10 15Met Gln Val Be Asp Asn Be Val Gly Leu Lys Trp Asn Glu Gln Met1 5 10 15

Gly Thr Thr Lys Lys Gly Tyr Thr Phe vai Gln Ala Asn Arg Ala Trp20 25 30Gly Thr Thr Lys Lys Gly Tyr Thr Phe Goes Gln Wing Asn Arg Wing Trp20 25 30

Leu Arg Lys Tyr Leu Leu Phe Trp Ile Met Gly Met Ala Phe Ile Ser35 40 45Leu Arg Lys Tyr Leu Leu Phe Trp Ile Met Gly Met Wing Phe Ile Ser35 40 45

Met Leu lie Tyr Asn Gly Met Asp Ala Asp Ile Lys vai Arg Arg Asn50 55 60Met Leu lie Tyr Asn Gly Met Asp Wing Asp Ile Lys will Arg Arg Asn50 55 60

Glu Val Leu Gly Ser Met Cys Glu Gln Arg Ala Arg Met Leu Gln Asp65 70 75 80Glu Val Leu Gly Ser Met Cys Glu Gln Arg Wing Arg Met Leu Gln Asp65 70 75 80

Gln Phe Asn Val Ser vai Asn His Val His Ala Leu Ala Val Leu Val85 90 95Gln Phe Asn Val Ser Will Asn His Val His Wing Leu Wing Val Leu Val85 90 95

Ser Thr Phe His Tyr Phe Lys Asn Pro Ser Ala Ile Asp Gln Glu Thr100 105 110Ser Thr Phe His Tyr Phe Lys Asn Pro Ser Ia Asp Asp Gln Glu Thr100 105 110

Phe Ala Glu Tyr Thr Ala Arg Thr Ala Phe Glu Arg Pro Leu Leu Ser115 120 125Phe Glu Wing Tyr Thr Arg Wing Wing Thr Phe Glu Arg Wing Wing Read Leu Ser115 120 125

Gly Val Ala Tyr Ala Gln Arg vai Ile His Ser Glu Arg Asp Ile Phe130 135 140Gly Val Wing Tyr Wing Gln Arg Goes Ile His Being Glu Arg Asp Ile Phe130 135 140

Glu Lys Gln His Gly Trp Met Ile Arg Thr Met Glu Lys Glu Pro Ser145 150 155 160Glu Lys His Gl Gly Trp Met Ile Arg Thr Met Glu Lys Glu Pro Ser145 150 155 160

Pro Asp Arg Asp Glu Tyr Ala Pro Val Ile Phe Ser Gln Glu Thr vaiPro Asp Arg Asp Glu Tyr Wing Pro Val Ile Phe Ser Gln Glu Thr Go

165 170 175165 170 175

Ser Tyr Ile Glu Ser Leu Asp Met Met Ser Gly Glu Glu Asp Arg Glu180 185 190Be Tyr Ile Glu Be Read Asp Met Met Be Gly Glu Glu Asp Arg Glu180 185 190

Asn Ile Leu Arg Ala Arg Ala Thr Gly Lys Ala vai Leu Thr Arg ProAsn Ile Leu Arg Wing Arg Wing Arg Wing Thr Gly Lys Wing Go Leu Thr Arg Pro

195 200 205195 200 205

Phe Arg Leu Leu Gly Ser His His Leu Gly vai vai Leu Thr Phe Pro210 215 220Phe Arg Leu Read Gly Be His His Leu Gly Goes Go Leu Thr Phe Pro210 215 220

Val Tyr Lys Phe Lys Leu Pro Ser Ile Pro Thr Glu Glu Glu Arg Ile225 230 235 240Val Tyr Lys Phe Lys Leu Pro Ser Ile Pro Thr Glu Glu Glu Arg Ile225 230 235 240

Glu Ala Thr Ala Gly Tyr Val Gly Gly Ala Phe Asp vai Glu Ser LeuGlu Wing Thr Wing Gly Tyr Val Gly Gly Wing Phe Asp Goes Glu Be Leu

245 250 255245 250 255

Val Glu Asn Leu Xaa Gly Gln Leu Ala Gly Asn Gln Ala Ile Leu Val260 265 270Val Glu Asn Leu Xaa Gly Gln Leu Wing Gly Asn Gln Wing Ile Leu Val260 265 270

Asn vai Tyr Asp vai Thr Asn Ser Ser Asp Leu Leu vai Met Tyr GlyAsn Will Tyr Asp Will Thr Asn Be Ser Asp Leu Leu Go Met Tyr Gly

275 280 285275 280 285

His Gln Tyr Gln Asp Gly Asp Leu ser Leu ser His Glu Ser Ser Leu290 295 300His Gln Tyr Gln Asp Gly Asp Read Be Read Read Be His Glu Be Read Leu290 295 300

Asp Phe Gly Asp Pro Phe Arg Lys His Leu Met Ile Cys Arg Tyr Gln305 310 315 320Asp Phe Gly Asp Pro Phe Arg Lys His Leu Met Ile Cys Arg Tyr Gln305 310 315 320

Gln Arg Ala Pro Thr ser Trp Thr Ala Leu Thr Thr Ala Phe Leu PheGln Arg Wing Thr Thr Be Wing Thr Thr Wing Thr Thr Wing Phe Leu Phe

325 330 335325 330 335

Phe Val Ile Gly Leu Leu Val Gly Tyr Ile Leu Tyr Gly Ala Ala Thr340 345 350Phe Val Ile Gly Leu Leu Val Gly Tyr Ile Leu Tyr Gly Wing Ward Thr340 345 350

His Ile Val Lys Val Glu Asp Asp Phe His Glu Met Gln Val Leu LysHis Ile Val Lys Val Glu Asp Asp Phe His Glu Met Gln Val Leu Lys

355 360 365355 360 365

vai Arg Ala Glu Ala Ala Asp Val Ala Lys Ser Gln Phe Leu Ala Thr370 375 380go Arg Wing Glu Wing Wing Asp Val Wing Lys Ser Gln Phe Leu Wing Thr370 375 380

val Ser His Glu lie Arg Thr Pro Met Asn Gly lie Leu Gly Met Leu385 390 395 400val Be His Glu lie Arg Thr Pro Met Asn Gly lie Leu Gly Met Leu385 390 395 400

Ala Leu Leu Leu Asp Thr Asp Leu Ser Ser Thr Gln Lys Asp Tyr Ala405 410 415Wing Read Leu Read Asp Thr Asp Read Be Ser Thr Gln Lys Asp Tyr Ala405 410 415

Gln Thr Ala Gln Ala Cys Gly Lys Ala Leu Ile Ala Leu Ile Asn Glu420 425 430Gln Thr Wing Gln Cys Wing Gly Lys Wing Leu Ile Wing Leu Ile Asn Glu420 425 430

Val Leu Asp Arg Ala Lys Ile Glu Ala Gly Lys Leu Glu Leu Glu Ala435 440 445Val Leu Asp Arg Wing Lys Ile Glu Wing Gly Lys Leu Glu Leu Glu Ala435 440 445

vai Pro Phe Asp Ile Arg Ser Ile Leu Asp Asp Val Leu ser Leu Phe450 455 460go to Phe Asp Ile Arg Be Ile Leu Asp Asp Val Leu Be Leu Phe450 455 460

Ser Glu Lys Ser Arg Gln Lys Gly Leu Glu Leu Ala vai Phe vai Ser465 470 475 480Be Glu Lys Be Arg Gln Lys Gly Leu Glu Leu Wing Go Phe Go Ser465 470 475 480

Asp Lys Val Pro Glu Ile Val Ile Gly Asp Pro Gly Arg Phe Arg Gln485 490 495Asp Lys Val Pro Glu Ile Val Ile Gly Asp Pro Gly Arg Phe Arg Gln485 490 495

Ile Ile Thr Asn Leu Val Gly Asn Ser Val Lys Phe Thr Glu Arg Gly500 505 510Ile Ile Thr Asn Read Val Gly Asn Ser Val Lys Phe Thr Glu Arg Gly500 505 510

His lie Phe Val Lys Val His Leu Ala Glu Asn Ser Lys Val Ser Met515 520 525His lie Phe Val Lys Val His Leu Wing Glu Asn Ser Lys Val Ser Met515 520 525

Asp Ser Glu Tyr vai Asn Gly Ile Ser Asp Ser Gly Leu Phe vai Leu530 535 540Asp Be Glu Tyr Will Asn Gly Ile Be Asp Be Gly Leu Phe Will Leu530 535 540

Asp Gly Arg Glu Phe Gln Thr Leu Ser Gly Arg Glu Ala Ala Asp Asp545 550 555 560Asp Gly Arg Glu Phe Gln Thr Read Ser Gly Arg Glu Wing Wing Asp Asp545 550 555 560

Gln Asn ser Trp Asp Asn Phe Lys His Leu Ile Ala Asp Asp Asn Phe565 570 575Gln Asn Be Trp Asp Asn Phe Lys His Leu Ile Wing Asp Asp Asn Phe565 570 575

Gln Ser Asn Ala Ala Ser Asn Asn Ser Ala Val Thr Asn Lys Gly Cys580 585 590Gln Be Asn Wing Ward Be Asn Asn Be Wing Val Thr Asn Lys Gly Cys580 585 590

Asp His vai Thr Leu Met vai Ser Val Glu Asp Thr Gly Ile Gly Ile595 600 605Asp His Will Thr Leu Met Will Be Val Glu Asp Thr Gly Ile Gly Ile595 600 605

Leu Leu His Ala Gln Asn Arg vai Phe Thr Pro Phe Met Gln Ala Asp610 615 620Leu Leu Read His Wing Gln Asn Arg Goes Phe Thr Pro Phe Met Gln Wing Asp610 615 620

Ser Ser Thr Ser Arg Asn Tyr Gly Gly Thr Gly Ile Gly Leu Ser Ile625 630 635 640Be Ser Thr Be Arg Asn Tyr Gly Gly Thr Gly Ile Gly Read Ser Be Ile625 630 635 640

Ser Lys Cys Leu Val Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Ser645 650 655Ser Lys Cys Leu Val Glu Leu Met Gly Gly Gln Ile Asn Phe Ile Ser645 650 655

Arg Pro Gln Ile Gly Ser Thr Phe Ser Phe Thr Ala Val Phe Gly Lys660 665 670Arg Pro Gln Ile Gly Be Thr Phe Be Phe Thr Wing Val Phe Gly Lys660 665 670

Cys Lys Lys Asn Ser Met Asn Asp Met Lys Lys Pro Asn Ser Glu GluCys Lys Lys Asn Be Met Asn Asp Met Lys Lys Pro Asn Be Glu Glu

675 680 685675 680 685

Leu Pro Pro Ser Phe Arg Gly Met Lys Ala Ile vai vai Asp Ser Lys5 690 695 700Leu Pro Pro Be Phe Arg Gly Met Lys Wing Ile Goes Go Asp Ser Lys5 690 695 700

His Val Arg Ala Ser Val Thr Arg Tyr His Leu Lys Arg Leu Gly Ile705 710 715 720His Val Arg Wing Be Val Thr Arg Tyr His Leu Lys Arg Leu Gly Ile705 710 715 720

Ile vai Glu Val Thr Asn Ser Ile Asn Met Ala Ala Ser Leu Phe Arg725 7B0 735Ile Go Glu Val Thr Asn Be Ile Asn Met Wing Wing Be Read Phe Arg725 7B0 735

10 Glu Asn Gly Ser Thr Leu Pro Arg Asn Thr Ile Leu Pro Asp Met Ile740 745 75010 Glu Asn Gly To Be Thr Read Le Arg Asn Thr Ile Le Le Pro Asp Met Ile740 745 750

Leu vai Glu Lys Asp Ile Leu Asn Ser Asp Glu Glu Cys Gly Ile IleLeu Goes Glu Lys Asp Ile Leu Asn Be Asp Glu Cys Gly Ile Ile

755 760 765755 760 765

His His Leu Asn Trp Lys Pro Asn Gly Ser Ser Val Lys Phe Pro Lys15 770 775 780His His Leu Asn Trp Lys Pro Asn Gly Ser Be Val Lys Phe Pro Lys15 770 775 780

Leu Ile Leu Leu Ala Thr Asn Ile Ala Thr Ala Glu Leu Asp Lys Ala785 790 795 800Leu Ile Leu Leu Wing Thr Thr Asn Ile Wing Thr Thr Glu Leu Asp Lys Ala785 790 795 800

Arg Ala Ala Gly Phe Ala Asp Thr vai Ile Met Lys Pro Leu Arg Ala805 810 815Arg Wing Gly Wing Phe Wing Asp Thr Go Ile Met Lys Pro Read Arg Wing Ala805 810 815

20 Thr Met Val Ala Ala Cys Leu Gln Gln vai Leu Gly Val Lys Asn Gln820 825 83020 Thr Met Val Wing Cys Wing Read Gln Gln Go Read Gly Val Lys Asn Gln820 825 830

Arg Arg Pro Asn Gly Ser Ala Phe Leu Gln Ser Leu Leu Cys Gly LysArg Arg Pro Asn Gly Be Ala Phe Leu Gln Be Leu Read Cys Gly Lys

835 840 845835 840 845

Arg Ile Leu Ile Val Asp Asp Asn Arg vai Asn Arg Arg vai Ala Ala25 850 855 860Arg Ile Leu Ile Val Asp Asp Asn Arg Go Asn Arg Arg Go Wing Ala25 850 855 860

Gly Ala Leu Lys Lys Phe Gly Ala Asp vai Glu Cys Ala Asp Ser Gly865 870 875 880Gly Wing Read Lys Lys Phe Gly Wing Asp Goes Glu Cys Wing Asp Ser Gly865 870 875 880

Lys Ser Ala Leu Lys Leu Leu Gln Leu Pro His Asn Phe Asp Ala Cys885 890 895Lys Ser Ala Leu Lys Leu Leu Gln Leu Pro His Asn Phe Asp Wing Cys885 890 895

30 Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Ala Thr Arg900 905 91030 Phe Met Asp Ile Gln Met Pro Glu Met Asp Gly Phe Glu Wing Thr Arg900 905 910

Arg Ile Arg Thr Met Glu Val Glu Ala Asn Lys Gly Gly Leu Ser Ala915 920Arg Ile Arg Thr Met Glu Val Glu Wing Asn Lys Gly Gly Leu Ser Ala915 920

Thr Glu Gly Lys Arg Pro Ile ProThr Glu Gly Lys Arg Pro Ile Pro

930 935930 935

Ile His Ala Thr Tyr Glu Glu Cys945 950Ile His Wing Thr Tyr Glu Glu Cys945 950

vai Ser Lys Pro Phe Glu Glu Glu965will be Lys Pro Phe Glu Glu Glu965

Phe Phe Lys Lys Pro980Phe Phe Lys Lys Pro980

925925

Ile Leu Ala Met Thr Ala Asp vai940Ile Leu Wing Met Thr Wing Asp vai940

Leu Lys Cys Gly Met Asn Gly Tyr955 960Read Lys Cys Gly Met Asn Gly Tyr955 960

Asn Leu Tyr Lys Glu Val Ala Arg970 975Asn Leu Tyr Lys Glu Val Wing Arg970 975

Claims (15)

1. Molécula de DNA que codifica um polipeptídeo de histidinaquinase mutante, caracterizada pelo fato de que compreende uma seqüên-cia de aminoácidos selecionada entre:a. SEQ ID NO: 6, 15 e 16, em que o resíduo de aminoácido 266[Phe] na SEQ ID NO: 6 e 266 [Xaa] na SEQ ID NO: 15 e resíduo 267 [Xaa]na SEQ ID NO: 16 é selecionado entre isoleucina, serina, treonina, valina,metionina, alanina, fenilalanina, tirosina, triptofano, arginina, lisina, glicina,histidina, aspartato, asparagina, glutamato, glutamina, prolina e cisteína,b. um truncamento de (a),em que o referido peptídeo é capaz de induzir formação espontânea quandoexpressa em uma planta.1. DNA molecule encoding a mutant histidine kinase polypeptide, characterized in that it comprises an amino acid sequence selected from: a. SEQ ID NO: 6, 15 and 16, wherein amino acid residue 266 [Phe] in SEQ ID NO: 6 and 266 [Xaa] in SEQ ID NO: 15 and residue 267 [Xaa] in SEQ ID NO: 16 is selected from isoleucine, serine, threonine, valine, methionine, alanine, phenylalanine, tyrosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine, proline and cysteine, b. a truncation of (a), wherein said peptide is capable of inducing spontaneous formation when expressed in a plant. 2. Polipeptídeo de histidina quinase mutante, caracterizado pelofato de ser codificado pela molécula de DNA tal como definida na reivindicação 1.Mutant histidine kinase polypeptide, characterized in that it is encoded by the DNA molecule as defined in claim 1. 3. Planta geneticamente modificada, caracterizada por ter umaseqüência de nucleotídeo que codifica um polipeptídeo de histidina quinasemutante que compreende uma seqüência de aminoácidos selecionada entre:a. SEQ ID NO: 6, 15 e 16, em que o resíduo de aminoácido 266[Phe]na SEQ ID NO: 6 e resíduo 266 [Xaa]na SEQ ID NO: 15 e resíduo 267[Xaa] na SEQ ID NO: 16 é selecionado entre isoleucina, serina, treonina,valina, metionina, alanina, fenilalanina, tirosina, triptofano, arginina, lisina,glicina, histidina, aspartato, asparagina, glutamato, glutamina, prolina e ciste-ína,b. um truncamento de (a),em que a dita planta é capaz de formação espontânea de nódulo.3. Genetically modified plant, characterized in that it has a nucleotide sequence encoding a kinasemutating histidine polypeptide comprising an amino acid sequence selected from: a. SEQ ID NO: 6, 15 and 16, wherein amino acid residue 266 [Phe] in SEQ ID NO: 6 and residue 266 [Xaa] in SEQ ID NO: 15 and residue 267 [Xaa] in SEQ ID NO: 16 is selected from isoleucine, serine, threonine, valine, methionine, alanine, phenylalanine, tyrosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine, proline and cysteine, b. a truncation of (a), wherein said plant is capable of spontaneous nodule formation. 4. Planta geneticamente modificada de acordo com a reivindi-cação 3, caracterizada pelo fato de que o polipeptídeo de histidina quinasemutante é codificado por uma molécula de ácido nucleico tendo uma se-quência de ácido nucleico selecionada do grupo: SEQ ID NO: 4, 5, e 7, ou aseqüência codificante dessas.Genetically modified plant according to claim 3, characterized in that the histine quinasemutant polypeptide is encoded by a nucleic acid molecule having a nucleic acid sequence selected from the group: SEQ ID NO: 4, 5, and 7, or the coding frequency thereof. 5. Uso de uma molécula de DNA que codifica um polipeptídeode histidina quinase mutante, caracterizado pelo fato de ser capaz de induzirformação de nódulo espontânea quando expressa em uma planta consistin-do em uma seqüência de aminoácido selecionado entre:а. SEQ ID NO: 6, 15 e 16, em que o resíduo de aminoácido 266[Phe]na SEQ ID NO: 6 e resíduo 266 [Xaa]na SEQ ID NO: 15 e resíduo 267[Xaa] na SEQ ID NO: 16 é selecionado entre isoleucina, serina, treonina,valina, metionina, alanina, fenilalanina, tirosina, triptofano, arginina, lisina,glicina, histidina, aspartato, asparagina, glutamato, glutamina, prolina e ciste-ína, eb. um truncamento de (a),como um transgene para produzir a planta geneticamente modificada comodefinida na reivindicação 5.5. Use of a DNA molecule encoding a mutant histidine kinase polypeptide, characterized in that it is capable of inducing spontaneous nodule formation when expressed in a plant consisting of an amino acid sequence selected from: а. SEQ ID NO: 6, 15 and 16, wherein amino acid residue 266 [Phe] in SEQ ID NO: 6 and residue 266 [Xaa] in SEQ ID NO: 15 and residue 267 [Xaa] in SEQ ID NO: 16 is selected from isoleucine, serine, threonine, valine, methionine, alanine, phenylalanine, tyrosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine, proline and cysteine, eb. a truncation of (a) as a transgene to produce the genetically modified plant as defined in claim 5. 6. Planta geneticamente modificada como definida na reivindi-cação 3 ou 4, caracterizada pelo fato de que compreende adicionalmenteuma seqüência de nucleotídeos homóloga ou heteróloga do promotor opera-velmente ligada à seqüência de nucleotídeos que codifica o polipeptídeo.Genetically modified plant as defined in claim 3 or 4, characterized in that it further comprises a promoter homologous or heterologous nucleotide sequence operably linked to the nucleotide sequence encoding the polypeptide. 7. Planta geneticamente modificada de acordo com a reivindi-cação 6, caracterizada pelo fato de que o dito promotor é um promotor regu-lado.Genetically modified plant according to claim 6, characterized in that said promoter is a regulated promoter. 8. Planta geneticamente modificada de acordo com qualqueruma das reivindicações 3, 4, 6 ou 7, caracterizada pelo fato de que a ditaplanta é uma planta monocotiledônea ou dicotiledônea.Genetically modified plant according to any one of claims 3, 4, 6 or 7, characterized in that the said plant is a monocotyledonous or dicotyledonous plant. 9. Planta geneticamente modificada de acordo com a reivindi-cação 8, caracterizada pelo fato de que a dita planta é selecionada entrearroz, cevada, milho, aveia, centeio, sorgo, trigo e grama Poaceae.Genetically modified plant according to claim 8, characterized in that said plant is selected from rice, barley, maize, oats, rye, sorghum, wheat and grass Poaceae. 10. Planta geneticamente modificada de acordo com qualqueruma das reivindicações 3, 4, 6 a 9, obtenível por um processo de mutagê-nese de DNA e seleção de uma planta capaz de formação espontânea denódulo, caracterizada pelo fato de ter uma seqüência de nucleotídeo quecodifica um polipeptídeo de histidina quinase mutante, compreendendo umaseqüência de aminoácido selecionada entre:a. SEQ ID NO: 6, 15 e 16, em que o resíduo de aminoácido 266[Phe]na SEQ ID NO: 6 e resíduo 266 [Xaa]na SEQ ID NO: 15 e resíduo 267[Xaa] na SEQ ID NO: 16 é selecionado entre isoleucina, serina, treonina,valina, metionina, alanina, fenilalanina, tirosina, triptofano, arginina, lisina,glicina, histidina, aspartato, asparagina, glutamato, glutamina, prolina e cisteína, eb. um truncamento de (a)Genetically modified plant according to any one of Claims 3, 4, 6 to 9, obtainable by a DNA mutagenesis process and selection of a plant capable of spontaneous formation in the nodule, characterized in that it has a nucleotide sequence which encodes a mutant histidine kinase polypeptide comprising an amino acid sequence selected from: a. SEQ ID NO: 6, 15 and 16, wherein amino acid residue 266 [Phe] in SEQ ID NO: 6 and residue 266 [Xaa] in SEQ ID NO: 15 and residue 267 [Xaa] in SEQ ID NO: 16 is selected from isoleucine, serine, threonine, valine, methionine, alanine, phenylalanine, tyrosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine, proline and cysteine, eb. a truncation of (a) 11. Método de produção de uma planta geneticamente modifi-cada tal como definida em qualquer uma das reivindicações 3, 4, 6 a 9, ca-racterizado pelo fato de que a introdução de cassete de gene compreen-dendo a dita seqüência de nucleotídeos que codifica o dito polipeptídeo eselecionando uma planta transgênica e a sua progênie que expressa o ditopolipeptídeo.A method of producing a genetically modified plant as defined in any one of claims 3, 4, 6 to 9, characterized in that the introduction of a gene cassette comprising said nucleotide sequence which encodes said polypeptide by selecting a transgenic plant and its progeny expressing the dithopolipeptide. 12. Método de acordo com a reivindicação 11, caracterizado pe-lo fato de que o cassete de gene é introduzido em uma planta pela trans-formação ou é sujeito à "introgress" na planta por cruzamento sexual com aplanta transgênica compreendendo o dito cassete de gene.A method according to claim 11, characterized in that the gene cassette is introduced into a plant by transformation or is subjected to "introgress" in the plant by transgenic plant sexual crossing comprising said cassette. gene. 13. Planta geneticamente modificada produzida de acordo como método como definido na reivindicação 11 ou 12, caracterizada pelo fatode ter uma seqüência de nucleotídeo que codifica um polipeptídeo de histi-dina quinase mutante, compreendendo uma seqüência de aminoácido sele-cionada entre:a. SEQ ID NO: 6, 15 e 16, em que o resíduo de aminoácido 266[Phe]na SEQ ID NO: 6 e resíduo 266 [Xaa]na SEQ ID NO: 15 e resíduo 267[Xaa] na SEQ ID NO: 16 é selecionado entre isoleucina, serina, treonina,valina, metionina, alanina, fenilalanina, tirosina, triptofano, arginina, lisina,glicina, histidina, aspartato, asparagina, glutamato, glutamina, prolina e ciste-ína, eb. um truncamento de (a)em que o referido peptídeo é capaz de induzir formação espontânea quandoexpressa em uma planta.Genetically modified plant produced according to a method as defined in claim 11 or 12, characterized in that it has a nucleotide sequence encoding a mutant histidine kinase polypeptide, comprising an amino acid sequence selected from: a. SEQ ID NO: 6, 15 and 16, wherein amino acid residue 266 [Phe] in SEQ ID NO: 6 and residue 266 [Xaa] in SEQ ID NO: 15 and residue 267 [Xaa] in SEQ ID NO: 16 is selected from isoleucine, serine, threonine, valine, methionine, alanine, phenylalanine, tyrosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine, proline and cysteine, eb. a truncation of (a) wherein said peptide is capable of inducing spontaneous formation when expressed in a plant. 14. Semente da planta geneticamente modificada tal como defi-nida em qualquer uma das reivindicações 3, 4, 6 a 10 e 13, caracterizadapelo fato de ter uma seqüência de nucleotídeo que codifica um polipeptídeode histidina quinase mutante, compreendendo uma seqüência de amino-ácido selecionada entre:a. SEQ ID NO: 6, 15 e 16, em que o resíduo de aminoácido 266[Phe]na SEQ ID NO: 6 e resíduo 266 [Xaa]na SEQ ID NO: 15 e resíduo 267[Xaa] na SEQ ID NO: 16 é selecionado entre isoleucina, serina, treonina,valina, metionina, alanina, fenilalanina, tirosina, triptofano, arginina, lisina,glicina, histidina, aspartato, asparagina, glutamato, glutamina, prolina e ciste-rna, eb. um truncamento de (a)em que o referido polipeptídeo é capaz de induzir formação de nódulo es-pontânea quando expressa em uma planta.Genetically modified plant seed as defined in any one of claims 3, 4, 6 to 10 and 13, characterized in that it has a nucleotide sequence encoding a mutant histidine kinase polypeptide comprising an amino acid sequence selected from: a. SEQ ID NO: 6, 15 and 16, wherein amino acid residue 266 [Phe] in SEQ ID NO: 6 and residue 266 [Xaa] in SEQ ID NO: 15 and residue 267 [Xaa] in SEQ ID NO: 16 is selected from isoleucine, serine, threonine, valine, methionine, alanine, phenylalanine, tyrosine, tryptophan, arginine, lysine, glycine, histidine, aspartate, asparagine, glutamate, glutamine, proline and cysteine, eb. a truncation of (a) wherein said polypeptide is capable of inducing spontaneous nodule formation when expressed in a plant. 15. Plantação, caracterizada pelo fato de que compreende umaplanta geneticamente modificada tal como definida em qualquer uma dasreivindicações 3, 4, 6 a 10 e 13.Planting, characterized in that it comprises a genetically modified plant as defined in any one of claims 3, 4, 6 to 10 and 13.